U.S. patent number 10,246,486 [Application Number 15/465,492] was granted by the patent office on 2019-04-02 for macrocyclic inhibitors of flaviviridae viruses.
This patent grant is currently assigned to Cypralis Limited, Gilead Sciences, Inc.. The grantee listed for this patent is Cypralis Limited, Gilead Sciences, Inc.. Invention is credited to Caroline Aciro, David Kenneth Dean, Neil Andrew Dunbar, Adrian John Highton, Petr Jansa, Kapil Kumar Karki, Andrew John Keats, Linos Lazarides, Richard Mackman, Simon Neil Pettit, Karine G. Poullennec, Adam James Schrier, Dustin Scott Siegel, Victoria Alexandra Steadman.
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United States Patent |
10,246,486 |
Aciro , et al. |
April 2, 2019 |
Macrocyclic inhibitors of flaviviridae viruses
Abstract
Provided are compounds of Formula I: ##STR00001## and
pharmaceutically acceptable salts and esters thereof. The
compounds, compositions, and methods provided are useful for the
treatment of virus infections, particularly hepatitis C
infections.
Inventors: |
Aciro; Caroline (Bottisham,
GB), Dean; David Kenneth (Ware, GB),
Dunbar; Neil Andrew (Chelmsford, GB), Highton; Adrian
John (Chelmsford, GB), Jansa; Petr (San Mateo,
CA), Karki; Kapil Kumar (Foster City, CA), Keats; Andrew
John (Essex, GB), Lazarides; Linos (London,
GB), Mackman; Richard (Millbrae, CA), Pettit;
Simon Neil (Essex, GB), Poullennec; Karine G.
(Essex, GB), Schrier; Adam James (Redwood City,
CA), Siegel; Dustin Scott (San Carlos, CA), Steadman;
Victoria Alexandra (Essex, GB) |
Applicant: |
Name |
City |
State |
Country |
Type |
Gilead Sciences, Inc.
Cypralis Limited |
Foster City
Cambridge |
CA
N/A |
US
GB |
|
|
Assignee: |
Gilead Sciences, Inc. (Foster
City, CA)
Cypralis Limited (Cambridge, GB)
|
Family
ID: |
48670106 |
Appl.
No.: |
15/465,492 |
Filed: |
March 21, 2017 |
Prior Publication Data
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Document
Identifier |
Publication Date |
|
US 20170190737 A1 |
Jul 6, 2017 |
|
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
14740680 |
Jun 16, 2015 |
9873716 |
|
|
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13913184 |
Jun 7, 2013 |
9090653 |
|
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61657553 |
Jun 8, 2012 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K
31/497 (20130101); C07K 5/06139 (20130101); C07K
5/06191 (20130101); C07D 498/22 (20130101); C07K
5/02 (20130101); C07D 471/18 (20130101); C07K
19/00 (20130101); A61P 31/12 (20180101); A61K
31/7072 (20130101); C07K 5/0606 (20130101); A61P
31/14 (20180101); A61K 38/21 (20130101); A61K
31/403 (20130101); C07K 5/06052 (20130101); A61K
31/7056 (20130101); A61P 1/16 (20180101); A61K
38/55 (20130101); A61K 39/29 (20130101); A61K
38/05 (20130101); A61P 31/20 (20180101); A61P
37/02 (20180101); C07D 487/08 (20130101); A61P
11/00 (20180101); A61P 31/18 (20180101); C07K
5/06078 (20130101); A61P 35/00 (20180101); A61P
29/00 (20180101); C07D 498/18 (20130101); C07D
498/08 (20130101); A61P 43/00 (20180101); A61K
45/06 (20130101); C07K 5/06034 (20130101); A61P
31/00 (20180101); C07K 5/06086 (20130101); A61K
31/504 (20130101); A61K 38/12 (20130101); A61P
3/04 (20180101); A61K 38/21 (20130101); A61K
2300/00 (20130101); A61K 38/55 (20130101); A61K
2300/00 (20130101); A61K 31/7056 (20130101); A61K
2300/00 (20130101); A61K 31/497 (20130101); A61K
2300/00 (20130101); A61K 31/403 (20130101); A61K
2300/00 (20130101); A61K 31/7072 (20130101); A61K
2300/00 (20130101); A61K 31/504 (20130101); A61K
2300/00 (20130101); A61K 38/00 (20130101); A61K
38/15 (20130101); Y02A 50/30 (20180101) |
Current International
Class: |
A61K
38/21 (20060101); A61K 31/403 (20060101); A61K
31/497 (20060101); A61K 31/504 (20060101); A61K
38/55 (20060101); A61K 31/7072 (20060101); A61K
31/7056 (20060101); C07D 498/22 (20060101); C07D
471/18 (20060101); C07K 5/02 (20060101); A61K
45/06 (20060101); A61K 39/29 (20060101); A61K
38/12 (20060101); C07D 498/18 (20060101); C07D
498/08 (20060101); C07D 487/08 (20060101); C07K
5/062 (20060101); C07K 19/00 (20060101); C07K
5/06 (20060101); C07K 5/068 (20060101); A61K
38/05 (20060101); C07K 5/078 (20060101); C07K
5/065 (20060101); A61K 38/00 (20060101); A61K
38/15 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
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1193979 |
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Sep 1998 |
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CN |
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CN |
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Apr 2008 |
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KR |
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Jan 1997 |
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WO |
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WO |
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WO |
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WO |
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WO |
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WO |
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WO-2011/098808 |
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WO |
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WO-2011/144924 |
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Nov 2011 |
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WO |
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WO-2012/040040 |
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Mar 2012 |
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WO |
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WO-2012/078915 |
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Jun 2012 |
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WO |
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WO-2012078915 |
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Jun 2012 |
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WO |
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WO-2013/185090 |
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Dec 2013 |
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WO |
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WO-2013/185093 |
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Dec 2013 |
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WO |
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WO-2013/185103 |
|
Dec 2013 |
|
WO |
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Primary Examiner: Bredefeld; Rachael E
Assistant Examiner: Yang; Kaipeen E
Attorney, Agent or Firm: Sheppard Mullin Richter &
Hampton LLP
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of U.S. patent application Ser.
No. 14/740,680, filed Jun. 16, 2015, now U.S. Pat. No. 9,873,716,
which is a divisional of U.S. patent application Ser. No.
13/913,184, filed Jun. 7, 2013, now U.S. Pat. No. 9,090,653, which
claims the benefit under 35 U.S.C. .sctn. 119(e) to U.S.
Provisional Application No. 61/657,553, filed Jun. 8, 2012. The
entirety of each application is incorporated herein by reference.
Claims
The invention claimed is:
1. A compound of Formula I: ##STR00385## or a pharmaceutically
acceptable salt, isotope, stereoisomer, mixture of stereoisomers,
tautomer, ester or prodrug thereof, wherein: A.sup.1 is
(C.sub.2-C.sub.5)alkylene, (C.sub.2-C.sub.5)alkenylene,
(C.sub.2-C.sub.5)alkynylene, --O--(C.sub.2-C.sub.4)alkylene,
--O--(C.sub.2-C.sub.4)alkenylene, arylene,
aryl(C.sub.1-C.sub.2)alkylene, heterocycloalkylene, pyrazolylene,
pyridylene, pyrimidinylene or
heterocycloalkyl(C.sub.1-C.sub.2)alkylene, wherein a sp.sup.3
carbon atom of A.sup.1 is optionally substituted with one or more
(C.sub.1-C.sub.4)alkyl; A.sup.2 is arylene or heteroarylene,
wherein A.sup.2 is optionally substituted with halo; X.sup.1 is
--O--, --NH-- or --N((C.sub.1-C.sub.4)alkyl)-; R.sup.1a and
R.sup.1b are independently H, (C.sub.1-C.sub.4)alkyl,
(C.sub.2-C.sub.4)alkenyl or (C.sub.2-C.sub.4)alkynyl; R.sup.2 is H,
(C.sub.1-C.sub.4)alkyl, (C.sub.2-C.sub.4)alkenyl or
(C.sub.2-C.sub.4)alkynyl; R.sup.3a and R.sup.3b are independently H
or (C.sub.1-C.sub.8)alkyl; R.sup.4a and R.sup.4b are independently
H, --OH, (C.sub.1-C.sub.4)alkoxy, halo(C.sub.1-C.sub.4)alkoxy or
(C.sub.1-C.sub.8)alkyl; R.sup.5 is H, (C.sub.1-C.sub.4)alkyl,
(C.sub.2-C.sub.4)alkenyl or (C.sub.2-C.sub.4)alkynyl, or R.sup.5
forms a cyclic moiety along with --N((C.sub.1-C.sub.4)alkyl)- of
X.sup.1 or arylene of A.sup.2; and R.sup.6 is H or
(C.sub.1-C.sub.4)alkyl.
2. The compound of claim 1, wherein A.sup.1 is ethenylene,
propenylene, butenylene, ethylene, propylene, butylene,
oxypropylene, oxypropenylene, pyrazolylene, phenylene, pyridylene
or pyrimidinylene.
3. The compound of claim 1, wherein A.sup.2 is isoquinolinylene,
phenylene or halophenylene.
4. The compound of claim 1, wherein X.sup.1 is --O-- or --NH--; one
of R.sup.1a and R.sup.1b is H and the other is methyl; R.sup.2 is
iso-propyl; R.sup.5 is methyl and R.sup.6 is H or methyl.
5. The compound of claim 1, wherein R.sup.3a is H or methyl;
R.sup.3b is H; R.sup.4a is H, --OH, methoxy, trifluoroethoxy; and
R.sup.4b is H.
6. The compound of claim 1, which is a compound of Formula II:
##STR00386## or a pharmaceutically acceptable salt, isotope,
stereoisomer, mixture of stereoisomers, tautomer, ester or prodrug
thereof, wherein: A.sup.1 is ethenylene, ##STR00387## A.sup.2 is
##STR00388## X.sup.1 is --O-- or --NH--; R.sup.3a is H or methyl;
R.sup.4a is H, --OH, (C.sub.1-C.sub.4)alkoxy,
halo(C.sub.1-C.sub.4)alkoxy or (C.sub.1-C.sub.8)alkyl; and R.sup.5
is H or (C.sub.1-C.sub.4)alkyl.
7. The compound of claim 1, wherein A.sup.2 is heteroarylene;
A.sup.1 is (C.sub.2-C.sub.5)alkylene, (C.sub.2-C.sub.5)alkenylene,
(C.sub.2-C.sub.5)alkynylene, wherein A.sup.1 is optionally
substituted with one or more (C.sub.1-C.sub.4)alkyl; R.sup.3a is H
or methyl; and R.sup.4a is H, --OH or (C.sub.1-C.sub.4)alkoxy.
8. The compound of claim 1, wherein A.sup.2 is arylene; and A.sup.1
is (C.sub.2-C.sub.5)alkylene, (C.sub.2-C.sub.5)alkenylene,
(C.sub.2-C.sub.5)alkynylene, O--(C.sub.2-C.sub.4)alkylene,
--O--(C.sub.2-C.sub.4)alkenylene, wherein A.sup.1 is optionally
substituted with one or more (C.sub.1-C.sub.4)alkyl; R.sup.3a is H
or methyl; and R.sup.4a is H, --OH, (C.sub.1-C.sub.4)alkoxy or
halo(C.sub.1-C.sub.4)alkoxy.
9. A compound selected from: ##STR00389## ##STR00390## ##STR00391##
##STR00392## or a pharmaceutically acceptable salt, isotope,
stereoisomer, mixture of stereoisomers, tautomer, ester or prodrug
thereof.
10. The compound of claim 1, wherein A.sup.2 is arylene; and
A.sup.1 is pyrazolylene, phenylene or pyridylene.
11. The compound of claim 1, wherein A.sup.2 is halophenylene; and
A.sup.1 is --O--(C.sub.2-C.sub.4)alkylene or
--O--(C.sub.2-C.sub.4)alkenylene.
12. The compound of claim 1, wherein A.sup.1 is
(C.sub.2-C.sub.5)alkylene or (C.sub.2-C.sub.4)alkenylene; R.sup.5
is methyl, or R.sup.5 form ##STR00393## along with arylene of
A.sup.2, or R.sup.5 form ##STR00394## along with
--N((C.sub.1-C.sub.4)alkyl)- of X.sup.1; and R.sup.6 is H or
methyl.
13. A compound selected from: ##STR00395## ##STR00396##
##STR00397## or a pharmaceutically acceptable salt, isotope,
stereoisomer, mixture of stereoisomers, tautomer, ester or prodrug
thereof.
14. A pharmaceutical composition comprising a compound of claim 1
or a pharmaceutically acceptable salt, isotope, stereoisomer,
mixture of stereoisomers, tautomer, ester or prodrug thereof and a
pharmaceutically acceptable excipient.
15. The pharmaceutical composition of claim 14, further comprising
at least one additional therapeutic agent selected from the group
consisting of interferons, ribavirin, HCV NS3 protease inhibitors,
HCV NS5a inhibitors, nucleoside or nucleotide inhibitors of HCV
NS5B polymerase, non-nucleoside inhibitors of HCV NS5B polymerase,
and TLR-7 agonists; or a mixture thereof.
16. The pharmaceutical composition of claim 15, wherein the at
least one additional therapeutic agent is ribavirin, telaprevir,
boceprevir or sofosbuvir.
Description
FIELD
The present application provides novel compounds inhibiting
viruses, compositions containing such compounds, and therapeutic
methods comprising the administration of such compounds.
BACKGROUND
RNA viruses comprising the Flaviviridae family include at least
three distinguishable genera including pestiviruses, flaviviruses,
and hepaciviruses (Calisher, et al., J. Gen. Virol., 1993, 70,
37-43). While pestiviruses cause many economically important animal
diseases such as bovine viral diarrhea virus (BVDV), classical
swine fever virus (CSFV, hog cholera) and border disease of sheep
(BDV), their importance in human disease is less well characterized
(Moennig, V., et al., Adv. Vir. Res. 1992, 48, 53-98). Flaviviruses
are responsible for important human diseases such as dengue fever
and yellow fever while hepaciviruses cause hepatitis C virus
infections in humans. Other important viral infections caused by
the Flaviviridae family include West Nile virus (WNV) Japanese
encephalitis virus (JEV), tick-borne encephalitis virus, Junjin
virus, Murray Valley encephalitis, St Louis enchaplitis, Omsk
hemorrhagic fever virus and Zika virus.
The hepatitis C virus (HCV) is the leading cause of chronic liver
disease worldwide (Boyer, N. et al. J Hepatol. 32:98-112, 2000) so
a significant focus of current antiviral research is directed
toward the development of improved methods of treatment of chronic
HCV infections in humans (Di Besceglie, A. M. and Bacon, B. R.,
Scientific American, October: 80-85, (1999); Gordon, C. P., et al.,
J. Med. Chem. 2005, 48, 1-20; Maradpour, D., et al., Nat. Rev.
Micro. 2007, 5(6), 453-463). A number of HCV treatments are
reviewed by Dymock et al. in Antiviral Chemistry &
Chemotherapy, 11:2; 79-95 (2000). Virologic cures of patients with
chronic HCV infection are difficult to achieve because of the
prodigious amount of daily virus production in chronically infected
patients and the high spontaneous mutability of HCV virus (Neumann,
et al., Science 1998, 282, 103-7; Fukimoto, et al., Hepatology,
1996, 24, 1351-4; Domingo, et al., Gene 1985, 40, 1-8; Martell, et
al., J. Virol. 1992, 66, 3225-9.
Currently, there are primarily two antiviral compounds, ribavirin,
a nucleoside analog, and interferon-alpha (a) (IFN), that are used
for the treatment of chronic HCV infections in humans. Ribavirin
alone is not effective in reducing viral RNA levels, has
significant toxicity, and is known to induce anemia. The
combination of IFN and ribavirin has been reported to be effective
in the management of chronic hepatitis C (Scott, L. J., et al.
Drugs 2002, 62, 507-556) but less than half the patients given this
treatment show a persistent benefit. Therefore, there is a need to
develop more effective anti-HCV therapies.
The macrocycle sanglifehrin and derivatives are immunomodulatory
and bind peptidyl-prolyl cis/trans isomerase (PPlase) cyclophilins
in a unique manner (WO 97/02285; WO 98/07743; J. Am. Chem. Soc
2003, 125, 3849-3859; J. Org. Chem. 2000, 65, 9255-9260; Angew.
Chem. Int. Ed. 1999, 38, 2443-2446). The cyclophilins are
peptidyl-prolyl cis/trans isomerases (PPlase) that regulate protein
folding in vivo and inhibit hepatitis C virus (Lin et al.,
WO2006/138507). However, none of the sanglifehrins or their
derivatives has become available for human anti-viral therapy.
Therefore, there is a continuing need to develop macrocyclic
sanglifehrins with anti-Flaviviridae virus activity and
particularly anti-HCV activity.
SUMMARY
In one embodiment, there is provided a compound represented by
Formula I:
##STR00002##
or a pharmaceutically acceptable salt, isotope, stereoisomer,
mixture of stereoisomers, tautomer, ester or prodrug thereof,
wherein:
A.sup.1 is (C.sub.2-C.sub.5)alkylene, (C.sub.2-C.sub.5)alkenylene,
(C.sub.2-C.sub.5)alkynylene, --O--(C.sub.2-C.sub.4)alkylene,
--O--(C.sub.2-C.sub.4)alkenylene, arylene,
aryl(C.sub.1-C.sub.2)alkylene, heterocycloalkylene or
heterocycloalkyl(C.sub.1-C.sub.2)alkylene, wherein a spa carbon
atom of A.sup.1 is optionally substituted with one or more
(C.sub.1-C.sub.4)alkyl;
A.sup.2 is arylene or heteroarylene, wherein A.sup.2 is optionally
substituted with halo;
X.sup.1 is --O--, --NH-- or --N((C.sub.1-C.sub.4)alkyl)-;
R.sup.1a and R.sup.1b are independently H, (C.sub.1-C.sub.4)alkyl,
(C.sub.2-C.sub.4)alkenyl or (C.sub.2-C.sub.4)alkynyl;
R.sup.2 is H, (C.sub.1-C.sub.4)alkyl, (C.sub.2-C.sub.4)alkenyl or
(C.sub.2-C.sub.4)alkynyl;
R.sup.3a and R.sup.3b are independently H or
(C.sub.1-C.sub.8)alkyl;
R.sup.4a and R.sup.4b are independently H, --OH,
(C.sub.1-C.sub.4)alkoxy, halo(C.sub.1-C.sub.4)alkoxy or
(C.sub.1-C.sub.8)alkyl;
R.sup.5 is H, (C.sub.1-C.sub.4)alkyl, (C.sub.2-C.sub.4)alkenyl or
(C.sub.2-C.sub.4)alkynyl, or
R.sup.5 forms a cyclic moiety along with
--N((C.sub.1-C.sub.4)alkyl)- of X.sup.1 or arylene of A.sup.2;
and
R.sup.6 is H or (C.sub.1-C.sub.4)alkyl.
In another embodiment, there is provided a pharmaceutical
composition comprising a compound of Formula I r a pharmaceutically
acceptable salt, isotope, stereoisomer, mixture of stereoisomers,
tautomer, ester or prodrug thereof and one or more pharmaceutically
acceptable carriers or excipients. In one aspect of the embodiment,
the pharmaceutical composition further comprises one or more
additional therapeutic agents.
In yet another embodiment, a method for treating Flaviviridae viral
infection is provided comprising administering a therapeutically
effective amount of a compound of Formula I r a pharmaceutically
acceptable salt, isotope, stereoisomer, mixture of stereoisomers,
tautomer, ester or prodrug thereof to a mammal in need thereof. In
one aspect of the embodiment, the treatment results in the
reduction of the in viral load or clearance of viral RNA in a
patient.
In yet another embodiment, a method for treating Coronaviridae
viral infection is provided comprising administering a
therapeutically effective amount of a compound of Formula I r a
pharmaceutically acceptable salt, isotope, stereoisomer, mixture of
stereoisomers, tautomer, ester or prodrug thereof to a mammal in
need thereof. In one aspect of the embodiment, the treatment
results in the reduction of the in viral load or clearance of viral
RNA in a patient.
DETAILED DESCRIPTION
Definitions
Unless stated otherwise, the following terms and phrases as used
herein are intended to have the following meanings:
"Alkanoyl" is RC(O)--; "alkanoyloxy" is RC(O)O--; and
"alkanoylamino" is RC(O)NR'--; where R is an alkyl group as defined
herein, and R' is hydrogen or alkyl.
"Alkenyl" refers to a straight or branched hydrocarbyl group with
at least one site of unsaturation, i.e., a carbon-carbon, sp.sup.2
double bond. In some embodiments, alkenyl is a C.sub.2-C.sub.20
alkenyl group, a C.sub.2-C.sub.10 alkenyl group or a
C.sub.2-C.sub.6 alkenyl group. Examples of alkenyl group include,
but are not limited to, vinyl (--CH.dbd.CH.sub.2), allyl
(--CH.sub.2CH.dbd.CH.sub.2), cyclopentenyl (--C.sub.5H.sub.7), and
5-hexenyl (--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.dbd.CH.sub.2).
"Alkenylene" refers to an unsaturated, branched or straight chain
or cyclic hydrocarbon radical having two monovalent radical centers
derived by the removal of two hydrogen atoms from the same or two
different carbon atoms of a parent alkene. For example, and
alkenylene group can have 2 to 20 carbon atoms, 2 to 10 carbon
atoms, or 2 to 6 carbon atoms. Typical alkenylene radicals include,
but are not limited to, 1,2-ethenylene (--CH.dbd.CH--).
"Alkoxy" is RO-- where R is alkyl, as defined herein. Non-limiting
examples of alkoxy groups include methoxy, ethoxy and propoxy.
"Alkyl" refers to a straight or branched chain hydrocarbyl group.
In an embodiment, alkyl has from 1 to 20 carbon atoms (i.e.,
C.sub.1-C.sub.20 alkyl). In some embodiments, alkyl is a
C.sub.1-C.sub.10 alkyl group or a C.sub.1-C.sub.6 alkyl group.
Examples of alkyl groups include, but are not limited to, methyl,
ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, hexyl,
heptyl, octyl, nonyl and decyl.
"Alkylene" refers to a saturated, branched or straight chain
radical or cyclic hydrocarbon radical having two monovalent radical
centers derived by the removal of two hydrogen atoms from the same
or two different carbon atoms of a parent alkane. For example, an
alkylene group can have 1 to 20 carbon atoms, 1 to 10 carbon atoms,
or 1 to 6 carbon atoms. Examples of alkylene radicals include, but
are not limited to, methylene (--CH.sub.2--), ethylene
(--CH.sub.2CH.sub.2--), propylene (--CH.sub.2CH.sub.2CH.sub.2--)
and butylene (--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--).
"Alkynyl" refers to a hydrocarbon containing normal, secondary or
tertiary carbon atoms with at least one site of unsaturation, i.e.,
a carbon-carbon, sp triple bond. For example, an alkynyl group can
have 2 to 20 carbon atoms (i.e., C.sub.2-C.sub.20 alkynyl), 2 to 12
carbon atoms (i.e., C.sub.2-C.sub.12 alkyne) or 2 to 6 carbon atoms
(i.e., C.sub.2-C.sub.6 alkynyl). Examples of alkynyl groups
include, but are not limited to, acetylenic (--CH) and propargyl
(--CH.sub.2C.ident.CH).
"Alkylamino" refers to an amino group substituted with one or more
alkyl groups. "Mono(alkyl)amino" or "(alkyl)amino" is RNH--, and
"di(alkyl)amino" or "(alkyl).sub.2amino" is R.sub.2N--, where each
of the R groups is alkyl as defined herein and are the same or
different. Examples of alkylamino groups include, but are not
limited to, methylamino, ethylamino, propylamino, butylamino,
dimethylamino, diethylamino and methylethylamno.
"Amino" refers to --NH.sub.2.
"Alkynylene" refers to an unsaturated, branched or straight chain
or cyclic hydrocarbon radical having two monovalent radical centers
derived by the removal of two hydrogen atoms from the same or two
different carbon atoms of a parent alkyne. For example, an
alkynylene group can have 2 to 20 carbon atoms, 2 to 10 carbon
atoms, or 2 to 6 carbon atoms. Typical alkynylene radicals include,
but are not limited to, acetylene (--C.ident.C--), propargylene
(--CH.sub.2C.ident.C--), and 4-pentynylene
(--CH.sub.2CH.sub.2CH.sub.2C.ident.C--).
"Aryl" refers to any monocyclic or bicyclic carbon ring of up to 7
atoms in each ring, wherein at least one ring is aromatic, or an
aromatic ring system of 5 to 14 carbons atoms which includes a
carbocyclic aromatic group fused with a 5- or 6-membered cycloalkyl
group. Examples of aryl groups include, but are not limited to,
phenyl, naphthyl, tetrahydronaphthyl and indanyl.
"Arylalkyl" refers to an alkyl as defined herein substituted with
an aryl radical.
"Arylene" refers to an aryl as defined above having two monovalent
radical centers derived by the removal of two hydrogen atoms from
two different carbon atoms of a parent aryl. Typical arylene
radicals include, but are not limited to, phenylene, e.g.,
##STR00003## and naphthylene, e.g.,
##STR00004##
"Arylalkylene" refers to an arylalkyl as defined above having two
monovalent radical centers derived by the removal of one hydrogen
atom from the aryl radical and the other hydrogen removed from the
alkyl radical of the group.
"Cycloalkyl" refers to a hydrocarbyl group containing at least one
saturated or partially unsaturated ring structure, and attached via
a ring carbon. Cycloalkyl groups include hydrocarbon mono-, bi-,
and poly-cyclic rings, whether fused, bridged, or spiro. In various
embodiments, it refers to a saturated or a partially unsaturated
C.sub.3-C.sub.12 cyclic moiety, examples of which include
cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl,
cyclohexenyl, cycloheptyl and cyclooctyl.
"Cycloalkylalkyl" refers to an alkyl moiety substituted with a
cycloalkyl group. Examples of cycloalkylalkyl groups include
cyclopropylmethyl, cyclobutylmethyl, cyclopentylethyl and
cyclohexylmethyl.
"Cycloalkylene" refers to a cycloalkyl, as defined herein, having
two monovalent radical centers derived by the removal of two
hydrogen atoms from the same or two different carbon atoms of a
parent cycloalkyl. Examples of cycloalkylene include, but are not
limited to, cyclopropylene, cyclobutylene, cyclopentylene and
cyclohexylene.
"Dialkylaminoalkyl" refers to an alkyl moiety substituted with a
dialkylamino group, wherein dialkylamino is as defined herein.
"Ester" means any ester of a compound in which any of the --COOH
functions of the molecule is replaced by a --C(O)OR function, or in
which any of the --OH functions of the molecule are replaced with a
--OC(O)R function, in which the R moiety of the ester is any
carbon-containing group which forms a stable ester moiety,
including but not limited to alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkylalkyl, aryl, arylalkyl, heterocyclyl, heterocyclylalkyl
and substituted derivatives thereof.
"Halo" refers to chloro (--Cl), bromo (--Br), fluoro (--F) or iodo
(--I).
"Haloalkoxy" refers to alkoxy, as defined herein, substituted with
one or more halo radicals.
"Haloalkoxyalkyl" refers to an alkyl moiety substituted with a
haloalkoxy group, as defined herein.
"Haloalkyl" refers to an alkyl group, in which one or more hydrogen
atoms of the alkyl group is replaced with a halogen atom. Examples
of haloalkyl groups include, but are not limited to, --CF.sub.3,
--CHF.sub.2, --CFH.sub.2 and --CH.sub.2CF.sub.3.
"Heterocycloalkyl" refers to a saturated or partially unsaturated
monocyclic, bicyclic or tricyclic group of 2 to 14 ring-carbon
atoms and, in addition to ring-carbon atoms, 1 to 4 heteroatoms
selected from P, N, O and S. The heterocyclic group can be attached
through a carbon atom or through a heteroatom, and when
substituted, the substituent can be bonded to a carbon atom or a
heteroatom. Examples of heterocyclyl include azetidinyl,
benzoimidazolyl, benzofuranyl, benzofurazanyl, benzopyrazolyl,
benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl,
carbolinyl, cinnolinyl, furanyl, imidazolyl, indolinyl, indolyl,
indolazinyl, indazolyl, isobenzofuranyl, isoindolyl, isoquinolyl,
isothiazolyl, isoxazolyl, naphthpyridinyl, oxadiazolyl, oxazolyl,
oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl,
pyridazinyl, pyridopyridinyl, pyridazinyl, pyridyl, pyrimidyl,
pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl, tetrahydropyranyl,
tetrahydrothiopyranyl, tetrahydroisoquinolinyl, tetrazolyl,
tetrazolopyridyl, thiadiazolyl, thiazolyl, thienyl, triazolyl,
azetidinyl, 1,4-dioxanyl, hexahydroazepinyl, piperazinyl,
piperidinyl, pyridin-2-onyl, pyrrolidinyl, morpholinyl,
thiomorpholinyl, dihydrobenzoimidazolyl, di hydrobenzofuranyl,
dihydrobenzothiophenyl, dihydrobenzoxazolyl, dihydrofuranyl,
dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl,
dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl,
dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl,
dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl,
dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl,
dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl,
methylenedioxybenzoyl, tetrahydrofuranyl, and tetrahydrothienyl and
N-oxides thereof.
"Heterocycloalkylene" refers to a heterocycloalkyl, as defined
above, having two monovalent radical centers derived by the removal
of two hydrogen atoms from the same or two different carbon atoms
of a parent heterocycloalkyl group.
"Heterocycloalkylalkylene" refers to an heterocycloalkyl as defined
above having two monovalent radical centers derived by the removal
of one hydrogen atom from the heterocycloalkyl radical and the
other hydrogen removed from the alkyl radical of the group.
"Heteroaryl" refers to a monocyclic, bicyclic or tricyclic ring
having up to 7 atoms in each ring, wherein at least one ring is
aromatic and contains from 1 to 4 heteroatoms in the ring selected
from the group consisting of N, O and S. Non-limiting examples of
heteroaryl include pyridyl, thienyl, furanyl, pyrimidyl,
imidazolyl, pyranyl, pyrazolyl, thiazolyl, thiadiazolyl,
isothiazolyl, oxazolyl, isoxazoyl, pyrrolyl, pyridazinyl,
pyrazinyl, quinolinyl, isoquinolinyl, benzofuranyl, dibenzofuranyl,
dibenzothiophenyl, benzothienyl, indolyl, benzothiazolyl,
benzooxazolyl, benzimidazolyl, isoindolyl, benzotriazolyl, purinyl,
thianaphthenyl and pyrazinyl. Attachment of heteroaryl can occur
via an aromatic ring, or, if heteroaryl is bicyclic or tricyclic
and one of the rings is not aromatic or contains no heteroatoms,
through a non-aromatic ring or a ring containing no heteroatoms.
"Heteroaryl" is also understood to include the N-oxide derivative
of any nitrogen containing heteroaryl.
"Heteroarylalkyl" refers to an alkyl group, as defined herein, in
which a hydrogen atom has been replaced with a heteroaryl
group.
"Heteroarylene" refers to a heteroaryl, as defined above, having
two monovalent radical centers derived by the removal of two
hydrogen atoms from the same or two different carbon atoms of a
parent heteroaryl group. Non-limiting examples of heteroarylene
groups are:
##STR00005##
"Hydroxyalkoxy" refers to an alkoxy, as defined herein, substituted
with a hydroxyl group (--OH). An example of hydroxyalkoxy is
hydroxyethoxy.
"Hydroxyalkyl" refers to an alkyl group substituted with at least
one hydroxy group. Examples of hydroxyalkyl groups include, but are
not limited to, hydroxymethyl, hydroxyethyl, hydroxypropyl and
hydroxybutyl.
"Prodrug" refers to any compound that when administered to a
biological system generates the drug substance, i.e., active
ingredient, as a result of spontaneous chemical reaction(s), enzyme
catalyzed chemical reaction(s), photolysis, and/or metabolic
chemical reaction(s). A prodrug is thus a covalently modified
analog or latent form of a therapeutically active compound.
Non-limiting examples of prodrugs include ester moieties,
quaternary ammonium moieties, glycol moieties, and the like.
The term "optionally substituted" refers to a moiety wherein all
substituents are hydrogen or wherein one or more of the hydrogens
of the moiety are replaced by non-hydrogen substituents. Multiple
substitutions on the same atom are also permitted where chemically
feasible (e.g., a dioxo substitution to provide --S(O).sub.2--,
geminal substituents, spiro cycloalkyl or heterocycloalkyl rings,
etc.). In some embodiments, "one or more" substituents is from one
to three substituents.
Embraced herein, where applicable, are permissible isomers such as
tautomers, racemates, enantiomers, diastereomers, atropisomers,
configurational isomers of double bonds (E- and/or Z-), cis- and
trans-configurations in ring substitution patterns, and isotopic
variants.
"Pharmaceutically acceptable" means suitable for use in
pharmaceutical preparations, generally considered as safe for such
use, officially approved by a regulatory agency of a national or
state government for such use, or being listed in the U. S.
Pharmacopoeia or other generally recognized pharmacopoeia for use
in animals, and more particularly in humans.
"Pharmaceutically acceptable carrier" refers to a diluent,
adjuvant, excipient, or carrier, or other ingredient which is
pharmaceutically acceptable and with which a compound of the
invention is administered.
"Pharmaceutically acceptable salt" refers to a salt which may
enhance desired pharmacological activity. Examples of
pharmaceutically-acceptable salts include acid addition salts
formed with inorganic or organic acids, metal salts and amine
salts. Examples of acid addition salts formed with inorganic acids
include salts with hydrochloric acid, hydrobromic acid, sulfuric
acid, nitric acid and phosphoric acid. Examples of acid addition
salts formed with organic acids such as acetic acid, propionic
acid, hexanoic acid, heptanoic acid, cyclopentanepropionic acid,
glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic
acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric
acid, benzoic acid, o-(4-hydroxy-benzoyl)-benzoic acid, cinnamic
acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,
1,2-ethanedisulfonic acid, 2-hydroxyethane-sulfonic acid,
benzenesulfonic acid, p-chlorobenzenesulfonic acid,
2-naphthalenesulfonic acid, p-toluenesulfonic acid, camphorsulfonic
acid, 4-methyl-bicyclo[2.2.2]oct-2-ene1-carboxylic acid,
gluco-heptonic acid, 4,4'-methylenebis(3-hydroxy-2-naphthoic) acid,
3-phenylpropionic acid, trimethyl-acetic acid, tertiary butylacetic
acid, lauryl sulfuric acid, gluconic acid, glutamic acid,
hydroxy-naphthoic acids, salicylic acid, stearic acid and muconic
acid. Examples of metal salts include salts with sodium, potassium,
calcium, magnesium, aluminum, iron, and zinc ions. Examples of
amine salts include salts with ammonia and organic nitrogenous
bases strong enough to form salts with carboxylic acids.
A compound of a given formula (e.g. the compound of Formula I,
which also includes Formula I-a, I-b, I-c, I-d, I-e or II) is
intended to encompass the compounds of the disclosure, and the
pharmaceutically acceptable salts, stereoisomers, mixture of
stereoisomers or tautomers of such compounds. Additionally, the
compounds of the disclosure may possess one or more asymmetric
centers, and can be produced as a racemic mixture or as individual
enantiomers or diastereoisomers. The number of stereoisomers
present in any given compound of a given formula depends upon the
number of asymmetric centers present (there are 2'' stereoisomers
possible where n is the number of asymmetric centers). The
individual stereoisomers may be obtained by resolving a racemic or
non-racemic mixture of an intermediate at some appropriate stage of
the synthesis or by resolution of the compound by conventional
means. The individual stereoisomers (including individual
enantiomers and diastereoisomers) as well as racemic and
non-racemic mixtures of stereoisomers are encompassed within the
scope of the present disclosure, all of which are intended to be
depicted by the structures of this specification unless otherwise
specifically indicated.
"Isomers" are different compounds that have the same molecular
formula. Isomers include stereoisomers, enantiomers and
diastereomers.
"Stereoisomers" are isomers that differ only in the way the atoms
are arranged in space. Stereoisomers include enantiomers and
diastereomers.
"Enantiomers" are a pair of stereoisomers that are
non-superimposable mirror images of each other. A 1:1 mixture of a
pair of enantiomers is a "racemic" mixture. The term "(.+-.)" is
used to designate a racemic mixture where appropriate.
"Diastereoisomers" are stereoisomers that have at least two
asymmetric atoms, but which are not mirror-images of each
other.
The absolute stereochemistry is specified according to the Cahn
Ingold Prelog R S system. When the compound is a pure enantiomer
the stereochemistry at each chiral carbon may be specified by
either R or S. Resolved compounds whose absolute configuration is
unknown are designated (+) or (-) depending on the direction
(dextro- or laevorotary) that they rotate the plane of polarized
light at the wavelength of the sodium D line.
Some of the compounds exist as tautomeric isomers or "tautomers".
Tautomeric isomers are in equilibrium with one another. For
example, amide containing compounds may exist in equilibrium with
imidic acid tautomers. Regardless of which tautomer is shown, and
regardless of the nature of the equilibrium among tautomers, the
compounds are understood by one of ordinary skill in the art to
comprise both amide and imidic acid tautomers. Thus, the amide
containing compounds are understood to include their imidic acid
tautomers. Likewise, the imidic acid containing compounds are
understood to include their amide tautomers.
Any formula or structure provided herein, including Formula I, I-a,
I-b, I-c, I-d, I-e or II, is also intended to represent unlabeled
forms as well as isotopically labeled forms of the compounds. An
"isotope" may have structures depicted by the formulas given herein
except that one or more atoms are replaced by an atom having a
selected atomic mass or mass number. Examples of isotopes that can
be incorporated into compounds of the disclosure include isotopes
of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and
chlorine, such as, but not limited to .sup.2H (deuterium, D),
.sup.3H (tritium), .sup.11C, .sup.13C, .sup.14C, .sup.15N,
.sup.18F, .sup.31P, .sup.32P, .sup.35S, .sup.36Cl and .sup.125I.
Various isotopically labeled compounds of the present disclosure,
for example those into which radioactive isotopes such as .sup.3H,
.sup.13C and .sup.14C are incorporated. Such isotopically labelled
compounds may be useful in metabolic studies, reaction kinetic
studies, detection or imaging techniques, such as positron emission
tomography (PET) or single-photon emission computed tomography
(SPECT) including drug or substrate tissue distribution assays or
in radioactive treatment of patients.
The disclosure also included compounds of Formula I, I-a, I-b, I-c,
I-d, I-e or II in which from 1 to n hydrogens attached to a carbon
atom is/are replaced by deuterium, in which n is the number of
hydrogens in the molecule. Such compounds exhibit increased
resistance to metabolism and are thus useful for increasing the
half life of any compound of Formul Formula I, I-a, I-b, I-c, I-d,
I-e or II when administered to a mammal. See, for example, Foster,
"Deuterium Isotope Effects in Studies of Drug Metabolism", Trends
Pharmacol. Sci. 5(12):524-527 (1984). Such compounds are
synthesized by means well known in the art, for example by
employing starting materials in which one or more hydrogens have
been replaced by deuterium.
Deuterium labelled or substituted therapeutic compounds of the
disclosure may have improved DMPK (drug metabolism and
pharmacokinetics) properties, relating to distribution, metabolism
and excretion (ADME). Substitution with heavier isotopes such as
deuterium may afford certain therapeutic advantages resulting from
greater metabolic stability, for example increased in vivo
half-life, reduced dosage requirements and/or an improvement in
therapeutic index. An .sup.18F labeled compound may be useful for
PET or SPECT studies. Isotopically labeled compounds of this
disclosure and prodrugs thereof can generally be prepared by
carrying out the procedures disclosed in the schemes or in the
examples and preparations described below by substituting a readily
available isotopically labeled reagent for a non-isotopically
labeled reagent. It is understood that deuterium in this context is
regarded as a substituent in the compound of Formula I, I-a, I-b,
I-c, I-d, I-e or II.
The concentration of such a heavier isotope, specifically
deuterium, may be defined by an isotopic enrichment factor. In the
compounds of this disclosure any atom not specifically designated
as a particular isotope is meant to represent any stable isotope of
that atom. Unless otherwise stated, when a position is designated
specifically as "H" or "hydrogen", the position is understood to
have hydrogen at its natural abundance isotopic composition.
Accordingly, in the compounds of this disclosure any atom
specifically designated as a deuterium (D) is meant to represent
deuterium.
"Therapeutically-effective amount" refers to an amount of a
compound that, when administered to a subject for treating a
disease, is sufficient to effect treatment for the disease.
"Therapeutically effective amount" can vary depending on the
compound, the disease and its severity, the age, the weight, etc.
of the subject to be treated.
The term "treating", and grammatical equivalents thereof, when used
in the context of treating a disease, means slowing or stopping the
progression of a disease, or ameliorating at least one symptom of a
disease, more preferably ameliorating more than one symptom of a
disease. For example, treatment of a hepatitis C virus infection
can include reducing the HCV viral load in an HCV infected human
being, and/or reducing the severity of jaundice present in an HCV
infected human being.
Compounds
The present application provides a compound represented by Formula
I:
##STR00006##
or a pharmaceutically acceptable salt, isotope, stereoisomer,
mixture of stereoisomers, tautomer, ester or prodrug thereof,
wherein:
A.sup.1 is (C.sub.2-C.sub.5)alkylene, (C.sub.2-C.sub.5)alkenylene,
(C.sub.2-C.sub.5)alkynylene, --O--(C.sub.2-C.sub.4)alkylene,
--O--(C.sub.2-C.sub.4)alkenylene, arylene,
aryl(C.sub.1-C.sub.2)alkylene, heterocycloalkylene or
heterocycloalkyl(C.sub.1-C.sub.2)alkylene, wherein a spa carbon
atom of A.sup.1 is optionally substituted with one or more
(C.sub.1-C.sub.4)alkyl;
A.sup.2 is arylene or heteroarylene, wherein A.sup.2 is optionally
substituted with halo;
X.sup.1 is --O--, --NH-- or --N((C.sub.1-C.sub.4)alkyl)-;
R.sup.1a and R.sup.1b are independently H, (C.sub.1-C.sub.4)alkyl,
(C.sub.2-C.sub.4)alkenyl or (C.sub.2-C.sub.4)alkynyl;
R.sup.2 is H, (C.sub.1-C.sub.4)alkyl, (C.sub.2-C.sub.4)alkenyl or
(C.sub.2-C.sub.4)alkynyl;
R.sup.3a and R.sup.3b are independently H or
(C.sub.1-C.sub.8)alkyl;
R.sup.4a and R.sup.4b are independently H, --OH,
(C.sub.1-C.sub.4)alkoxy, halo(C.sub.1-C.sub.4)alkoxy or
(C.sub.1-C.sub.8)alkyl;
R.sup.5 is H, (C.sub.1-C.sub.4)alkyl, (C.sub.2-C.sub.4)alkenyl or
(C.sub.2-C.sub.4)alkynyl, or
R.sup.5 forms a cyclic moiety along with
--N((C.sub.1-C.sub.4)alkyl)- of X.sup.1 or arylene of A.sup.2;
and
R.sup.6 is H or (C.sub.1-C.sub.4)alkyl.
In certain embodiments, provided is a compound of Formula I:
##STR00007##
or a pharmaceutically acceptable salt, ester or prodrug thereof,
wherein:
A.sup.1 is (C.sub.1-C.sub.5)alkylene, (C.sub.2-C.sub.5)alkenylene,
(C.sub.2-C.sub.5)alkynylene, --O--(C.sub.2-C.sub.4)alkylene,
--O--(C.sub.2-C.sub.4)alkenylene, arylene,
aryl(C.sub.1-C.sub.2)alkylene, heterocycloalkylene or
heterocycloalkyl(C.sub.1-C.sub.2)alkylene, wherein a spa carbon
atom of A.sup.1 is optionally substituted with one or more
(C.sub.1-C.sub.4)alkyl;
A.sup.2 is arylene or heteroarylene, wherein A.sup.2 is optionally
substituted with halo;
X.sup.1 is --O--, --NH-- or --N((C.sub.1-C.sub.4)alkyl)-;
R.sup.1a and R.sup.1b are independently H, (C.sub.1-C.sub.4)alkyl,
(C.sub.2-C.sub.4)alkenyl or (C.sub.2-C.sub.4)alkynyl;
R.sup.2 is H, (C.sub.1-C.sub.4)alkyl, (C.sub.2-C.sub.4)alkenyl or
(C.sub.2-C.sub.4)alkynyl;
R.sup.3a and R.sup.3b are independently H or
(C.sub.1-C.sub.8)alkyl;
R.sup.4a and R.sup.4b are independently H, --OH,
(C.sub.1-C.sub.4)alkoxy, halo(C.sub.1-C.sub.4)alkoxy or
(C.sub.1-C.sub.8)alkyl;
R.sup.5 is H, (C.sub.1-C.sub.4)alkyl, (C.sub.2-C.sub.4)alkenyl or
(C.sub.2-C.sub.4)alkynyl, or
R.sup.5 forms a cyclic moiety along with
--N((C.sub.1-C.sub.4)alkyl)- of X.sup.1 or arylene of A.sup.2;
and
R.sup.6 is H or (C.sub.1-C.sub.4)alkyl.
In one aspect of the embodiment, A.sup.1 is ethenylene,
propenylene, butenylene, ethylene, propylene, butylene,
oxypropylene, oxypropenylene, pyrazolylene, phenylene or
pyrimidinylene.
In another aspect of the embodiment, A.sup.2 is heteroarylene such
as isoquinolinylene, phenylene or halophenylene. In one aspect,
A.sup.2 is
##STR00008##
In another aspect of the embodiment, X.sup.1 is --O-- or --NH--;
R.sup.1a is H; Rib is methyl; R.sup.2 is iso-propyl; R.sup.5 is
methyl and R.sup.6 is H or methyl.
In another aspect of the embodiment, R.sup.3a is H or methyl;
R.sup.3b is H; R.sup.4a is H, --OH, methoxy, trifluoroethoxy; and
R.sup.4b is H.
In another aspect of the embodiment, A.sup.2 is heteroarylene;
A.sup.1 is (C.sub.2-C.sub.5)alkylene, (C.sub.2-C.sub.5)alkenylene
or (C.sub.2-C.sub.5)alkynylene, wherein A.sup.1 is optionally
substituted with one or more (C.sub.1-C.sub.4)alkyl; R.sup.3a is H
or (C.sub.1-C.sub.8)alkyl; and R.sup.4a is H, --OH or
(C.sub.1-C.sub.4)alkoxy. In another aspect of the embodiment,
A.sup.2 is heteroarylene; A.sup.1 is (C.sub.1-C.sub.5)alkylene,
(C.sub.2-C.sub.5)alkenylene or (C.sub.2-C.sub.5)alkynylene, wherein
A.sup.1 is optionally substituted with one or more
(C.sub.1-C.sub.4)alkyl; R.sup.3a is H or (C.sub.1-C.sub.8)alkyl;
and R.sup.4a is H, --OH or (C.sub.1-C.sub.4)alkoxy. Non-limiting
examples of such compounds include the following compounds and
pharmaceutically acceptable salts thereof.
##STR00009##
TABLE-US-00001 Compound No. X.sup.1 A.sup.1 R.sup.3a R.sup.4a a-1
--NH-- --CH.sub.2CH.sub.2-- methyl methoxy a-2 --NH-- --CH.dbd.CH--
methyl methoxy a-3 --O-- --CH.sub.2CH.sub.2-- methyl methoxy a-4
--O-- --CH.dbd.CH-- methyl methoxy a-5 --NH-- --CH.sub.2CH.sub.2--
methyl --H a-6 --NH-- --CH.dbd.CH-- methyl --H a-7 --O--
--CH.sub.2CH.sub.2-- methyl --H a-8 --O-- --CH.dbd.CH-- methyl --H
a-9 --NH-- --CH.dbd.CHCH.sub.2-- --H --H a-10 --NH--
--CH.sub.2CH.sub.2CH.sub.2-- --H --H a-11 --O--
--CH.dbd.CHCH.sub.2-- --H --H a-12 --O--
--CH.sub.2CH.sub.2CH.sub.2-- --H --H a-13 --NH--
--CH.dbd.CHCH.sub.2CH.sub.2-- --H --H a-14 --NH--
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- --H --H a-15 --O--
--CH.dbd.CHCH.sub.2CH.sub.2-- --H --H a-16 --O--
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- --H --H
wherein the left bond of A.sup.1 linker is attached to A.sup.2.
In another aspect of the embodiment, A.sup.2 is arylene; and
A.sup.1 is (C.sub.2-C.sub.5)alkylene, (C.sub.2-C.sub.5)alkenylene,
(C.sub.2-C.sub.5)alkynylene, --O--(C.sub.2-C.sub.5)alkylene or
--O--(C.sub.2-C.sub.4)alkenylene, wherein A.sup.1 is optionally
substituted with one or more (C.sub.1-C.sub.4)alkyl; R.sup.3a is H
or (C.sub.1-C.sub.4)alkyl; and R.sup.4a is H, --OH,
(C.sub.1-C.sub.4)alkoxy or halo(C.sub.1-C.sub.4)alkoxy. In another
aspect of the embodiment, A.sup.2 is arylene; and A.sup.1 is
(C.sub.1-C.sub.5)alkylene, (C.sub.2-C.sub.5)alkenylene,
(C.sub.2-C.sub.5)alkynylene, --O--(C.sub.2-C.sub.5)alkylene or
--O--(C.sub.2-C.sub.4)alkenylene, wherein A.sup.1 is optionally
substituted with one or more (C.sub.1-C.sub.4)alkyl; R.sup.3a is H
or (C.sub.1-C.sub.4)alkyl; and R.sup.4a is H, --OH,
(C.sub.1-C.sub.4)alkoxy or halo(C.sub.1-C.sub.4)alkoxy.
Non-limiting examples of such compounds include the following
compounds and pharmaceutically acceptable salts thereof.
##STR00010##
TABLE-US-00002 No. X.sup.1 A.sup.1 R.sup.1b R.sup.3a R.sup.4a
R.sup.5 b-1 --NH-- --CH.dbd.CHCH.sub.2CH.sub.2-- --H --H --OH
methyl b-2 --NH-- --CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- --H --H --OH
methyl b-3 --O-- --CH.dbd.CHCH.sub.2CH.sub.2-- --H --H --OH methyl
b-4 --O-- --CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- --H --H --OH methyl
b-5 --NH-- --CH.dbd.CHCH.sub.2CH.sub.2-- --H --H
--OCH.sub.2CF.sub.3 methy- l b-6 --NH--
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- --H --H --OCH.sub.2CF.sub.- 3
methyl b-7 --O-- --CH.dbd.CHCH.sub.2CH.sub.2-- --H --H
--OCH.sub.2CF.sub.3 methyl- b-8 --O--
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- --H --H --OCH.sub.2CF.sub.3-
methyl b-9 --NH-- --CH.dbd.CHCH.sub.2C(CH.sub.3).sub.2-- --H --H
--H methyl b-10 --O-- --CH.dbd.CHCH.sub.2C(CH.sub.3).sub.2-- --H
--H --H methyl b-11 --NH-- --OCH.sub.2CH.dbd.CH-- --H methyl
methoxy isopropyl b-12 --O-- --OCH.sub.2CH.dbd.CH-- --H methyl
methoxy isopropyl b-13 --NH-- --OCH.sub.2CH.dbd.CH-- --H methyl
methoxy methyl b-14 --O-- --OCH.sub.2CH.dbd.CH-- --H methyl methoxy
methyl b-15 --NH-- --CH.dbd.CHCH.sub.2CH.sub.2-- methyl --H --OH
methyl b-16 --NH-- --CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- methyl --H
--OH methyl
wherein the left bond of A.sup.1 linker is attached to A.sup.2.
Also included are the following compounds and pharmaceutically
acceptable salts thereof.
##STR00011##
TABLE-US-00003 No. X.sup.1 A.sup.1 R.sup.3a R.sup.4a c-1 --NH--
--CH.dbd.CHCH.sub.2CH.sub.2-- methyl methoxy c-2 --NH--
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- methyl methoxy c-3 --O--
--CH.dbd.CHCH.sub.2CH.sub.2-- methyl methoxy c-4 --O--
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- methyl methoxy c-5 --NH--
--CH.dbd.CHCH.sub.2CH.sub.2-- --H --OH c-6 --NH--
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- --H --OH c-7 --O--
--CH.dbd.CHCH.sub.2CH.sub.2-- --H --OH c-8 --O--
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- --H --OH
wherein the left bond of A.sup.1 linker is attached to A.sup.2.
In another aspect of the embodiment, A.sup.2 is phenylene; and
A.sup.1 is pyrazolylene, phenylene or pyrimidinylene. Non-limiting
examples of such compounds include the following compounds and
pharmaceutically acceptable salts thereof.
##STR00012##
TABLE-US-00004 Compound No. X.sup.1 A.sup.1 R.sup.3a R.sup.4a d-1
--O-- ##STR00013## --H --H d-2 --O-- ##STR00014## --H --H d-3 --O--
##STR00015## --H --H d-4 --O-- ##STR00016## --H --H d-5 --NH--
##STR00017## --H --H d-6 --NH-- ##STR00018## --H --H d-7 --NH--
##STR00019## --H --H d-8 --NH-- ##STR00020## --H --H
wherein the left bond of A.sup.1 linker is attached to A.sup.2.
In another aspect of the embodiment, A.sup.2 is halophenylene; and
A.sup.1 is --O--(C.sub.2-C.sub.5)alkylene or
--O--(C.sub.2-C.sub.4)alkenylene. Non-limiting examples of such
compounds include the following compounds and pharmaceutically
acceptable salts thereof.
##STR00021##
TABLE-US-00005 Compound No. X.sup.1 A.sup.1 R.sup.3a R.sup.4a e-1
--NH-- --OCH.sub.2CH.dbd.CH-- methyl --H e-2 --O--
--OCH.sub.2CH.dbd.CH-- methyl --H e-3 --NH--
--OCH.sub.2CH.sub.2CH.sub.2-- methyl --H e-4 --O--
--OCH.sub.2CH.sub.2CH.sub.2-- methyl --H e-5 --NH--
--OCH.sub.2CH.dbd.CH-- methyl methoxy e-6 --O--
--OCH.sub.2CH.dbd.CH-- methyl methoxy e-7 --NH--
--OCH.sub.2CH.sub.2CH.sub.2-- methyl methoxy e-8 --O--
--OCH.sub.2CH.sub.2CH.sub.2-- methyl methoxy
wherein the left bond of A.sup.1 linker is attached to A.sup.2.
In one aspect of the embodiment, A.sup.1 is
(C.sub.2-C.sub.5)alkylene or (C.sub.2-C.sub.5)alkenylene; R.sup.5
is methyl, or R.sup.5 form
##STR00022## along with arylene of A.sup.2, or R.sup.5 form
##STR00023## along with --N((C.sub.1-C.sub.4)alkyl)- of X.sup.1;
and R.sup.6 is H or methyl. In another aspect of the embodiment,
A.sup.1 is (C.sub.1-C.sub.5)alkylene or
(C.sub.2-C.sub.5)alkenylene; R.sup.5 is methyl, or R.sup.5 form
##STR00024## along with arylene of A.sup.2, or R.sup.5 form
##STR00025## along with --N((C.sub.1-C.sub.4)alkyl)- of X.sup.1;
and R.sup.6 is H or methyl. Non-limiting examples of such compounds
include the following compounds and pharmaceutically acceptable
salts thereof:
##STR00026## ##STR00027##
In another embodiment, there is provided a compound of Formula
II:
##STR00028##
or a pharmaceutically acceptable salt, isotope, stereoisomer,
mixture of stereoisomers, tautomer, ester or prodrug thereof,
wherein:
A.sup.1 is ethenylene,
##STR00029## wherein the left bond of A.sup.1 linker is attached to
A.sup.2;
A.sup.2 is
##STR00030##
X.sup.1 is --O-- or --NH--;
R.sup.3a is H or (C.sub.1-C.sub.4)alkyl;
R.sup.4a is H, --OH, (C.sub.1-C.sub.4)alkoxy,
halo(C.sub.1-C.sub.4)alkoxy or (C.sub.1-C.sub.8)alkyl; and
R.sup.5 is H or (C.sub.1-C.sub.4)alkyl.
One skilled in the art will recognize that substituents and other
moieties of the compounds of the generic formulae herein should be
selected in order to provide a compound which is sufficiently
stable to provide a pharmaceutically useful compound which can be
formulated into an acceptably stable pharmaceutical composition.
Compounds which have such stability are contemplated as falling
within the scope of the present invention. It should be understood
by one skilled in the art that any combination of the definitions
and substituents described above should not result in an inoperable
species or compound.
Preparation of Macrocyclic Compounds
A compound of the present invention such as those of Formula I,
I-a, I-b, I-c, I-d, I-e or II can be prepared according to the
schemes described below, but it shall be appreciated that
modifications of the illustrated process or other process can also
be used. As illustrated in Scheme 1, the macrocyclic compounds M
are synthesized from the five key components A-E by combining them
together in sequence with the appropriate use of protecting groups
(PG.sup.1-PG.sup.8) by one skilled in the art. The hashed lines
numbered 1-5, hereby referred to as Connection 1, Connection 2,
etc., respectively, are the 5 connections for combining Components
A-E. The order in which the specific connections occur, can vary,
and are dependent on the choice of protecting groups and chemistry
required. In certain embodiments, Connections 3, 4 or 5 are used as
the final macrocyclization step.
##STR00031##
Illustratively, Connections 1 through 5 can be performed as
described below:
Connections 1, 2 and 3 are amide bonds. The connections are made
between the respective acid and amine using standard peptide
coupling agents (EDC/HOBT, DCC, PyBOP, PyBROP, HATU, HBTU, COMO,
etc.) known to one skilled in the art. The acid and amine coupling
partners are combined with a coupling agent in an organic solvent,
e.g., DMF, dichloromethane, acetonitrile, etc., in the presence of
a base, e.g., DIPEA, triethylamine, etc., at RT or slightly
elevated temperature. When any of these three steps are chosen as
the final macrocyclization step, for example Connection 3, then
macrolactamization conditions are preferred. Suitable
macrolactamization procedures include, but are not limited to,
those found in the following reference: Davies, J. S. J. Peptide
Sci. 2003, 9, 471-501.
Connection 4 is typically a carbon-carbon bond or a
heteroatom-carbon bond where the heteroatom is O, S or N. When
Connection 4 is a carbon-carbon bond, then standard carbon-carbon
bond forming procedures typically involving metal mediated cross
coupling reactions are preferred. Preferably the carbon-carbon bond
is formed using a Heck type coupling between an sp2 halide group
and an terminal alkene, a Suzuki coupling between an sp2 halide
group and a vinyl or aryl boronate, or a ring closing metathesis
(RCM) between two alkenes. Stille reactions can also be performed
between a vinyl stannane and an aryl or vinyl halide as described
in Journal of American Chemical Society 2000, 122, 3830 Nicolaou et
al. In each of the examples above the aryl or vinyl halide group
can also be an aryl or vinyl triflate.
For example, when A.sup.2-Y.sup.2 in A contains a terminal alkene,
such as --CH.dbd.CH.sub.2, and A.sup.1-Y.sup.1 in B contains a
terminal alkene, or CH.sub.3--CH.dbd.CH-- then a cross metathesis
reaction is performed. The two components are mixed in solvent,
e.g., acetonitrile, toluene and a metathesis catalyst, e.g., Grubbs
I, Grubbs II or Hoyveda-Grubbs I, Hoyveda-Grubbs II is added
followed by heating. If this connection is the final procedure to
close the macrocyclic ring, RCM conditions are preferred (e.g.,
more dilute conditions to avoid dimerization). For relevant RCM
conditions and examples see Journal of American Chemical Society
2003, 125, 3849 Sedrani et al and Journal of American Chemical
Society 2000, 122, 3830 Nicolaou et al. A typical RCM procedure
includes heating (either conventionally or by microwave) of the
acyclic precursor in a solvent such as toluene, or
1,2-dichloroethane, in the presence of a RCM catalyst, e.g., Grubbs
I, Grubbs II or Hoyveda-Grubbs I, Hoyveda-Grubbs II.
Alternatively, when Connection 4 is made via a Heck coupling
reaction, the vinyl or aryl halide, or the triflate A and the
alkene component B are mixed in a polar solvent, e.g.,
acetonitrile, or toluene in the presence of a Palladium(II)
catalyst, e.g., Palladium(OAc).sub.2, a phosphine ligand, e.g.,
P(o-toluene).sub.3, P(t-butyl).sub.3, etc., and a base, e.g.,
triethylamine. The reaction mixture is heated either conventionally
or in a microwave reactor.
Alternatively, when Connection 4 is made via a Suzuki coupling
reaction, the vinyl or aryl halide, or the triflate A and the vinyl
or aryl boronate B are mixed in a suitable solvent, e.g.,
cyclopentyl methyl ether, toluene, DMF, DME, etc., in the presence
of a Palladium catalyst (e.g.,
Palladium(II)Cl.sub.2(p-NMe.sub.2Ph).sub.2 and K.sub.3PO.sub.4 or
tetrakis(triphenylphosphine)palladium(0) and a base, such as
potassium carbonate). The reaction mixture is heated either
conventionally or in a microwave reactor. It is also possible in
such a coupling reaction to reverse the reactive functionalities on
the two starting materials, such that A is an aryl or vinyl
boronate and B contains a vinyl or aryl halide or triflate.
Alternatively, Connection 4 can be a carbon-oxygen bond and in this
case typical alkylation or nucleophilic aromatic substitution
conditions can be used between a hydroxyl group and an alkyl
halide, or aryl (or heteroaryl) halide. The hydroxyl reagent is
mixed with the alkyl or heteroaryl halide (preferably an iodide or
bromide), in an inert solvent, e.g., CPME, DMF, THF, etc, in the
presence base, e.g., cesium carbonate, cesium hydroxide, sodium
hydride, MaHMDS, etc.; and heated.
Alternatively, Connection 4 can be a carbon-nitrogen bond and in
this case typical alkylation, nucleophilic aromatic substitution or
Buchwald conditions can be used between an amine group and an alkyl
halide or heteroaryl halide. For example, the amine and the alkyl
or heteroaryl halide are mixed and heated in an inert solvent,
e.g., CPME, in the presence base, e.g., cesium carbonate, sodium
hydride, etc. An alternative procedure for the carbon-nitrogen
connection is to perform a reductive amination between an amine and
a carbonyl compound. Typically the amine and aldehyde or ketone are
mixed in an inert solvent, e.g., THF, dioxane and treated after a
period of time with sodium acetoxy borohydride or alternative
reducing agent.
Connection 5 is typically an amide (X.sup.1.dbd.--NH or substituted
N), or ester (X.sup.1.dbd.O) bond. When forming the amide bond,
standard coupling procedures as described for Connections 1-3 can
be used. In some embodiments, this is the final step in closing the
macrocycle. As such, macrolactamization types of coupling
procedures are more effective. Suitable macrolactamization
procedures include, but are not limited to, those found in Davies,
J. S. J. Peptide Sci. 2003, 9, 471-501.
When forming the ester bond, standard coupling reagents (e.g., EDC,
DCC, PyBOP, HATU, COMO) can be used, or when this is the final step
in formation of the macrocycle, macrolactonization procedures are
preferred (e.g., Shina, Yamaguchi). An exemplary method for the
macrolactonization step can be found in Journal of American
Chemical Society 2002, 124, 4257 Paquette et al or Chemical Reviews
2006, 106(3), 911-939. Typically, the acid and alcohol are mixed in
a polar solvent, e.g., DMF, acetonitrile, etc, in the presence of
the coupling agent and a base, e.g., DIPEA, DMAP.
The following general schemes provide general examples and
sequences for constructing the macrocyclic compound M from the
common precursors A-E.
##STR00032##
Compounds A-E can be deprotected (PG.sup.2-PG.sup.8) using
conditions described in Greene and Wuts, Protective Groups in
Organic Synthesis, John Wley and Sons, Inc. to provide Compounds
1a-1f.
In many cases the optimal protecting groups and their deprotection
methods are as follows. For Compound E, the typical protecting
group PG.sup.1 for the acid is a methyl or trichloroethyl ester.
The methyl and trichloroethyl esters can be removed by base, e.g.,
LiOH in a polar solvent, e.g., aqueous THF, etc. The trichloroethyl
ester can also be removed by treatment with zinc and ammonium
acetate in a polar solvent, e.g., THF. Typically, PG.sup.2 and
PG.sup.4 are acid labile groups, e.g., BOC, and are deprotected
using HCl in dioxane, or TMSOTf in dioxane, dichloromethane.
Typically, PG.sup.3 and PG.sup.5 are ester groups, removed by
treatment with alkali metal hydroxide in aqueous THF or dioxane.
Typically PG.sup.6 is an acid labile group, e.g., BOC, for an amine
and removed as described for PG.sup.2, or a silyl ether for a
hydroxyl group, and can be removed by treatment with HF.pyridine of
TBAF in an organic solvent, e.g., dichloromethane. Typically
PG.sup.8 is an amine protecting group, e.g., BOC, and is removed as
described for PG.sup.2 or a silyl ether for a hydroxyl group, which
can be removed as described for PG.sup.6, or an acetate protecting
group, which can be removed with an alkali metal hydroxide in
aqueous THF or dioxane.
Compound 1f is then coupled to acid 1a using the conditions
described above for Connection 1 to produce compound 1g. Compound
1g is then deprotected using conditions described in Greene and
Wuts and coupled to 1c to provide 1h using the conditions described
above for Connection 2. An alternative sequence for generating 1h
begins with coupling amine 1b to acid 1c using conditions as
described for Connection 2 above, to form 1i; deprotection of the
protecting group PG.sup.3 in 1i using conditions described in
Greene and Wuts, and finally coupling with amine 1f using
conditions described for Connection 1 above forms 1h.
##STR00033##
Compound 1h is deprotected at PG.sup.6 using conditions described
in Greene and Wuts and the amine is then coupled to 1d using the
conditions described above for Connection 3 to form 1j. Protecting
group PG.sup.1 in Compound 1j is then removed using conditions
described in Greene and Wuts, and the acid is then coupled to 1e
using conditions described for Connection 5 to form the acyclic
intermediate 1k. An alternative sequence to 1k is first
deprotection of PG.sup.1 and then coupling to 1e as described for
Connection 5; followed by deprotection of PG.sup.6 as described in
Greene and Wuts, followed by coupling to 1d using the conditions
described for Connection 3 to form 1k. Acyclic intermediate 1k is
then subjected to the RCM as described above for Connection 4 to
form the macrocycle M.
##STR00034##
Compound A is coupled to Compound B using the conditions described
above for Connection 4 to generate 2a. Compound 2a is then
deprotected at PG.sup.7 as described in Greene and Wuts to generate
acid 2b. Acid 2b is then coupled to the deprotected product of 1 h
(prepared from 1h by deprotection of PG.sup.6 described in Greene
and Wuts) to generate the precursor 2c. Deprotection of 2c is
carried out using conditions described in Greene and Wuts, and then
the product is cyclized using the conditions described above for
macrolactamization or macrolactonization in Connection 5, to
provide Compound M.
##STR00035##
Compound 2a is deprotected at PG.sup.8 as described in Greene and
Wuts and above in Connection 5 to generate 3a which is then
coupled, using conditions described above for Connection 5, to the
deprotected product of 1 h (prepared from 1 h by deprotection of
PG.sup.1 described in Greene and Wuts) to generate the precursor
3b. Deprotection of 3b is carried out using conditions described in
Greene and Wuts, and then cyclized using the conditions described
above for macrolactamization or macrolactonization in Connection 3,
to provide Compound M.
##STR00036##
For example, when M contains a C.dbd.C as a result of RCM, Compound
M is mixed in a solvent such as ethanol, methanol, etc., in the
presence of palladium on carbon catalyst under an atmosphere of
hydrogen gas to provide reduced Compound M1. The final macrocycle M
from Schemes 4-6 often contains protecting groups on side-chains
that require further removal to generate the final compound M.
Protecting groups on the R.sup.4a, R.sup.4b, R.sup.3a, R.sup.3b,
A.sup.1, A.sup.2 and/or X.sup.1 are removed using conditions
described in Greene and Wuts to generate Compound M2. Another
transformation is click chemistry to produce triazole M3. This
transformation is performed by treating the alkyne or azide in M,
in solvent (e.g., DMF) with an alkyne or azide as appropriate in
the presence of CuI to form M3.
Deprotected compound M2 can be further transformed after
deprotection to additional macrocycle M. For example, treatment of
M2 containing an --OH with an alkyl halide in the presence of a
base, e.g., cesium carbonate, in a suitable solvent, e.g., DMF,
acetonitrile, etc., forms alkylated product M4. M2 containing a
ketone group is treated in a suitable solvent (e.g., DMF, methanol,
etc.) with an amine followed by the addition of sodium
acetoxyborohydride to form amine product M5.
##STR00037##
Many B components containing an acid or ester with a terminal
alkene or CH.sub.3--CH.dbd.C-- or vinyl/aryl boronate groups are
commercially available or are described in the literature and can
be used in the above schemes directly. In addition, the following
schemes are examples of methods that can be used to generate
additional B components.
##STR00038## ##STR00039## ##STR00040##
In Scheme 8, part 1, a protected acid is treated with a strong
deprotonating base, e.g., LDA in an inert solvent, e.g., THF, at
-78.degree. C. and HMPA. A pre-cooled solution of dichloroacteylene
(prepared by treatment of trichlorethene with potassium hydride and
MeOH (catalytic) in THF) is then added to generate the chloro
acetylene product. This product is then reduced, for example, by
treatment with Cu in acetic acid and THF, to generate the alkyne,
which is then further reduced to the alkene, for example, by
treatment of an alcoholic solution of the alkyne with a poisoned
palladium reducing agent (e.g., Lindlar) in the presence of
hydrogen gas. Alternatively, the alkyne is treated with
Cp.sub.2ZrHCl in dichloromethane in the presence of pinnacolborane
to form the vinyl boronate.
In Scheme 8, part 2, a beta-keto ester with alpha substitution is
converted to the vinyl triflate, for example, by treating a THF
solution of the beta-keto ester with a base, e.g., LDA, in THF at
-78.degree. C., followed by addition of PhN(Tf).sub.2. The triflate
product is then treated with pyridine at elevated temperature to
form the alkyne. The alkyne is then treated as described above in
part 1 to generate the alkene or vinyl boronate products.
In Scheme 8, part 3, a chiral aldol reaction is used. An acyl group
is first attached to a chiral auxiliary, e.g., Evans, Oppolzer
sultam (see JACS 1990, 112, p 2767), using the standard amide bond
formation conditions as described above for Connection 1-3. The
Oppolzer auxiliary product is treated with the aldehyde of choice,
TBDMSOTf and base, e.g., triethylamine, in anhydrous solvent, e.g.,
dichloromethane. The Evans auxiliary is treated with base, e.g.,
LDA, KHMDS, DIPEA, in organic solvent, e.g., THF, at -78.degree. C.
and the aldehyde of choice in the presence of a Lewis acid, e.g.,
TiCl.sub.4, SnCl.sub.4, BF.sub.3OEt.sub.2. Protection of the
resulting alcohol from the aldol reaction is performed as described
in Greene and Wuts, or alternatively alkylation with an alkyl
halide or Meerwein's reagent, i.e., treatment with trimethyloxonium
tetrafluoroborate in an inert solvent, e.g., dichloromethane, is
performed. The auxiliary is then removed using standard alkali
metal hydroxide removal conditions, e.g., LiOH in THF, or LiOH and
hydrogen peroxide in THF, to provide the free acids product.
In Scheme 8, part 4, an Evans auxiliary is allylated with an allyl
halide as described in Synlett 2002, 12, 2039-2040. The product is
then isomerized by treatment with RhCl.sub.3 in ethanol and then
the auxiliary removed by base and peroxide, e.g., LiOH and
H.sub.2O.sub.2 in THF/Water. Alternatively the auxiliary is
directly removed by LiOH and H.sub.2O.sub.2 in THF/Water to provide
the terminal alkene.
In Scheme 8, part 5, a Homer Wadsworth Emmons reaction is used on
an aldehyde (containing a terminal alkene) to generate the
alpha-beta unsaturated ester, which is then selectively reduced to
the ester. For example, the phosphonate is treated with base, e.g.,
sodium hydride, in THF at low temperature, followed by addition of
the aldehyde and warming to generate the unsaturated ester. The
product is reduced by treatment with magnesium powder in
methanol.
In Scheme 8, part 6, an alpha-beta unsaturated acid is converted to
the unsaturated Evans auxiliary (see Organic Letters 2007, 9, p
1635) and is treated with an aldehyde to generate the corresponding
alkene product. The hydroxyl group is then protected using methods
described in Greene and Wuts and then the auxiliary is removed by
treatment with base and peroxide, e.g., LiOH and H.sub.2O.sub.2 in
THF/Water. The hydroxyl can also be alkylated by as described above
for aldol Scheme 8, part 3.
In Scheme 8, part 7, a ketone is transformed via the nitro olefin
as described in Angew. Chem. Int. Ed. 2006, 45 (46), 7736. The
nitro olefin is then treated with vinyl magnesium bromide in an
inert solvent, e.g., THF, in the presence of a copper(I) salt,
e.g., CuI and trimethylsilyl chloride. The nitro alkyl product
after addition of the vinyl group is then converted to the acid by
treating with sodium nitrite and acetic acid in an inert polar
solvent, e.g., DMSO.
Several types of A are available commercially or described in the
literature where X.sup.1 is O or NH and Y.sup.2 is a halide or
A.sup.2-Y.sup.2 contains an alkene. The schemes below describe
additional general methods for generating A.
##STR00041##
In Scheme 9, part 1 (Y.sup.2 is a halogen in A); the starting
Compound 6a is typically a commercially available aromatic
compound, that contains halogen Y.sup.2 and a group Z that can be
transformed to the ketone 6b. Typical Z groups are halide, acid, or
aldehyde, for example.
When Z is an acid, 6a is treated with a coupling agent, e.g., HATU,
EDC in the presence of a base, e.g., DIPEA and the Weinreb amine
(Me-NH--OMe) to form the Weinreb amide. The amide is then treated
with a nucleophile, e.g., TMS-CF.sub.3 to form the CF.sub.3
substituted ketone 6b or with a Grignard agent, e.g., MeMgBr in a
solvent, e.g., THF, at -78.degree. C. to form the methyl ketone
6b.
When Z is a halogen, then the initial conversion, if required, to a
more reactive halogen is performed by treatment with NaI and acetyl
chloride in an inert solvent, e.g., acetonitrile. The halogen is
then transformed to the ketone by a Stille reaction with an
ethoxyvinyl stannane. The halide is treated in an inert solvent,
e.g., toluene, with the stannane and a palladium (II) catalyst,
e.g., PdCl.sub.2(PPh.sub.3).sub.2, followed by treatment of the
product with 2M HCl to afford ketone 6b. In some cases the
formation of an alkyl lithium reagent from the halide group can be
performed by treatment with nBuLi at -78.degree. C. in THF and then
addition of a N-methoxy-N-methyl amide to afford the ketone 6b
(e.g., N-methoxy-N-methylacetamide affords the R.sup.5 is methyl
ketone 6b). A final method to generate the ketone 6b is through a
vinyl group. 6a is treated with a vinyltrifluoroborate in the
presence of a palladium catalyst, e.g., PdCl.sub.2(dppf) and then
the vinyl product is subsequently ozonolysed in a polar solvent,
e.g., methanol at low temperature to give an aldehyde. The aldehyde
is then reacted with a nucleophile, e.g., TSM-CF.sub.3 or a
Grignard reagent, e.g., MeMgBr to afford a secondary alcohol
product. The secondary alcohol is then oxidized with Dess Martin
Periodinane to give the desired ketone 6b or can be used as A
itself.
Chiral alcohol (X.sup.1 is O) and amine (X.sup.1 is NH) A are
generated using Chiral reduction methods on the ketone 6b. Chiral
alcohol 6c is formed from 6b using one of the numerous chiral
reduction methods available in the literature. Typically,
dichloro(p-cumene)ruthenium(II) dimer and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine are combined in
water, and sodium formate and 6b is added in a water miscible
solvent such as tetrahydrofuran. The reaction is then stirred at a
temperature between ambient and reflux to produce 6c where X.sup.1
is O. Alternatively, a chiral CBS reduction can be performed in an
inert solvent, e.g., THF at low temperature to also afford the
chiral alcohol 6c. Protection of the OH in 6c is performed using
methods described in Greene and Wuts, typically a TBS ether or
acetyl group are used to provide A (X.sup.1 is O).
Alternatively, to make chiral A (X.sup.1 is NH), ketone 6b is first
converted to a chiral imine (R.sup.10 is chiral group) and then
reduced using a variety of methods described in the literature. For
example, a chiral sulfinamide is reacted with the ketone 6b to
afford a chiral sulfinimine 6d, which is then reduced with a
suitable reducing agent, typically NaBH.sub.4, or selectride, or a
Noyori type reduction as described for the chiral alcohol above,
with dichloro(p-cumene)ruthenium(II) dimer and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine. The sulfinamide
auxiliary is then removed by treatment with mineral acid,
preferably HCl in a suitable organic solvent such as methanol, to
afford 6e where X.sup.1 is NH. Protection of the NH group can then
be performed as described in Greene and Wuts to generate A (X.sup.1
is NH).
In part 2 of Scheme 9, the synthesis of compound A where Y.sup.2 is
H and A.sup.2-Y.sup.2 contains --CH.dbd.CH.sub.2, a precursor for
metathesis and cross coupling reactions is illustrated. Exemplary
methods are as follows.
Compound 6c or 6e generated in Scheme 9, part 1 is first optionally
protected on X.sup.1 using a suitable protecting group as described
in Greene and Wuts, and then a vinyl group is introduced by a
suitable cross coupling method onto the aryl or sp2 halide. For
example, a transition metal mediated coupling with a vinyl stannane
or vinyl tetrafluoroborate using a suitable palladium catalyst,
e.g., PdCl.sub.2(dppf).sub.2 or PdCl.sub.2(PPh.sub.3).sub.2 in a
suitable organic solvent, e.g., acetonitrile, dichloromethane,
etc., with either thermal or microwave heating affords alkene
A.
Another typical method that can be used to introduce a vinyl group
is starting from the ketone 6b where Y is OH. Initially, triflation
of the alcohol is performed by treatment with Tf.sub.2O in the
presence of a base, e.g., pyridine. The ketone group is then
reduced with a Noyori reduction, or as described above through the
sulfonamide, to provide the chiral alcohol or amine. The chiral
alcohol or amine is then protected as described in Greene and Wuts,
and then the triflate is reacted with a vinyl cross coupling
reagent, e.g., vinyl stannane in a Stille coupling, or a
vinyltrifluoroborate as described above to introduce the alkene. A
further example of alkene generation using 6b ketone is via
introduction of an allyl group. Thus, 6b where Y is OH is treated
in an inert solvent in the presence of a suitable base, e.g.,
alkali metal carbonate, preferably potassium carbonate with allyl
bromide to form 6f. Compound 6f is then similar to ketone 6b and is
therefore able to be transformed as described above in part 1 to
Compound A where X.sup.1 is O or NH with protecting group
PG.sup.8.
Pharmaceutical Formulations
The compounds of this invention are formulated with conventional
carriers and excipients, which will be selected in accord with
ordinary practice. Tablets will contain excipients, glidants,
fillers, binders and the like. Aqueous formulations are prepared in
sterile form, and when intended for delivery by other than oral
administration generally will be isotonic.
While it is possible for the active ingredients to be administered
alone it may be preferable to present them as pharmaceutical
formulations. The formulations of the invention, both for
veterinary and for human use, comprise at least one active
ingredient, together with one or more acceptable carriers and
optionally other therapeutic ingredients.
Formulations of the present invention suitable for oral
administration may be presented as discrete units such as capsules,
cachets or tablets each containing a predetermined amount of the
active ingredient; as a powder or granules; as a solution or a
suspension in an aqueous or non-aqueous liquid; or as an
oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The
active ingredient may also be administered as a bolus, electuary or
paste.
The effective dose of an active ingredient depends at least on the
nature of the condition being treated, toxicity, whether the
compound is being used prophylactically (lower doses) or against an
active viral infection, the method of delivery, and the
pharmaceutical formulation, and will be determined by the clinician
using conventional dose escalation studies. The effective dose can
be expected to be from about 0.0001 to about 100 mg/kg body weight
per day; typically, from about 0.01 to about 10 mg/kg body weight
per day; more typically, from about 0.01 to about 5 mg/kg body
weight per day; most typically, from about 0.05 to about 0.5 mg/kg
body weight per day. For example, the daily candidate dose for an
adult human of approximately 70 kg body weight will range from 1 mg
to 1000 mg, preferably between 5 mg and 500 mg, and may take the
form of single or multiple doses.
Combination Therapy
The compounds of the present invention may be combined with one or
more active agents. Non-limiting examples of suitable active agents
to be combined include one or more interferons, ribavirin or its
analogs, HCV NS3 protease inhibitors, NS5a inhibitors,
alpha-glucosidase 1 inhibitors, hepatoprotectants, mevalonate
decarboxylase antagonists, antagonists of the renin-angiotensin
system, other anti-fibrotic agents, endothelin antagonists,
nucleoside or nucleotide inhibitors of HCV NSSB polymerase,
non-nucleoside inhibitors of HCV NSSB polymerase, HCV NSSA
inhibitors, TLR-7 agonists, cyclophillin inhibitors, HCV IRES
inhibitors, pharmacokinetic enhancers and other drugs for treating
HCV; or mixtures thereof.
More specifically, one or more compounds to be combined are
selected from the group consisting of
1) interferons, e.g., pegylated rIFN-alpha 2b (PEG-Intron),
pegylated rIFN-alpha 2a (Pegasys), rIFN-alpha 2b (Intron A),
rIFN-alpha 2a (Roferon-A), interferon alpha (MOR-22, OPC-18,
Alfaferone, Alfanative, Multiferon, subalin), interferon alfacon-1
(Infergen), interferon alpha-n1 (Wellferon), interferon alpha-n3
(Alferon), interferon-beta (Avonex, DL-8234), interferon-omega
(omega DUROS, Biomed 510), albinterferon alpha-2b (Albuferon), IFN
alpha XL, BLX-883 (Locteron), DA-3021, glycosylated interferon
alpha-2b (AVI-005), PEG-Infergen, PEGylated interferon lambda
(PEGylated IL-29), and belerofon;
2) ribavirin and its analogs, e.g., ribavirin (Rebetol, Copegus),
and taribavirin (Virami3);
3) HCV NS3 protease inhibitors, e.g., boceprevir (SCH-503034,
SCH-7), telaprevir (VX-950), VX-813, TMC-435 (TMC435350), ABT-450,
BI-201335, BI-1230, MK-7009, SCH-900518, VBY-376, VX-500, GS-9256,
GS-9451, BMS-790052, BMS-605339, PHX-1766, AS-101, YH-5258, YH5530,
YH5531, and ITMN-191 (R-7227);
4) alpha-glucosidase 1 inhibitors, e.g., celgosivir (MX-3253),
Miglitol, and UT-231B;
5) hepatoprotectants, e.g., emericasan (IDN-6556), ME-3738, GS-9450
(LB-84451), silibilin, and MitoQ;
6) nucleoside or nucleotide inhibitors of HCV NS5B polymerase,
e.g., R1626, R7128 (R4048), IDX184, IDX-102, PSI-7851, BCX-4678,
valopicitabine (NM-283), GS-6620 and MK-0608;
7) non-nucleoside inhibitors of HCV NS5B polymerase, e.g.,
filibuvir (PF-868554), ABT-333, ABT-072, BI-207127, VCH-759,
VCH-916, JTK-652, MK-3281, VBY-708, VCH-222, A848837, ANA-598,
GL60667, GL59728, A-63890, A-48773, A-48547, BC-2329, VCH-796
(nesbuvir), GSK625433, BILN-1941, XTL-2125, and GS-9190;
8) HCV NSSA inhibitors, e.g., AZD-2836 (A-831), AZD-7295 (A-689),
and BMS-790052;
9) TLR-7 agonists, e.g., imiquimod, 852A, GS-9524, ANA-773,
ANA-975, AZD-8848 (DSP-3025), PF-04878691, and SM-360320;
10) cyclophillin inhibitors, e.g., DEBIO-025, SCY-635, and
NIM811;
11) HCV IRES inhibitors, e.g., MCI-067;
12) pharmacokinetic enhancers, e.g., BAS-100, SPI-452, PF-4194477,
TMC-41629, GS-9350, GS-9585, and roxythromycin;
13) other drugs for treating HCV, e.g., thymosin alpha 1 (Zadaxin),
nitazoxanide (Alinea, NTZ), BIVN-401 (virostat), PYN-17 (altirex),
KPE02003002, actilon (CPG-10101), GS-9525, KRN-7000, civacir,
GI-5005, XTL-6865, BIT225, PTX-111, ITX2865, TT-033i, ANA 971,
NOc-205, tarvacin, EHC-18, VGX-410C, EMZ-702, AVI 4065, BMS-650032,
BMS-791325, Bavituximab, MDX-1106 (ONO-4538), Oglufanide, FK-788,
and VX-497 (merimepodib);
14) mevalonate decarboxylase antagonists, e.g., statins, HMGCoA
synthase inhibitors (e.g., hymeglusin), squalene synthesis
inhibitors (e.g., zaragozic acid);
15) angiotensin II receptor antagonists, e.g., losartan,
irbesartan, olmesartan, candesartan, valsartan, telmisartan,
eprosartan;
16) angiotensin-converting enzyme inhibitors, e.g., captopril,
zofenopril, enalapril, ramipril, quinapril, perindopril,
lisinopril, benazepril, fosinopril;
17) other anti-fibrotic agents, e.g., amiloride; and
18) endothelin antagonists, e.g. bosentan and ambrisentan.
In yet another embodiment, the present application provides a
combination therapy comprising a composition of the present
invention and a second pharmaceutical composition comprising at
least one additional therapeutic agent selected from the group
consisting of HIV protease inhibiting compounds, HIV non-nucleoside
inhibitors of reverse transcriptase, HIV nucleoside inhibitors of
reverse transcriptase, HIV nucleotide inhibitors of reverse
transcriptase, HIV integrase inhibitors, gp41 inhibitors, CXCR4
inhibitors, gp120 inhibitors, CCR5 inhibitors, interferons,
ribavirin analogs, NS3 protease inhibitors, NS5a inhibitors,
alpha-glucosidase 1 inhibitors, cyclophilin inhibitors,
hepatoprotectants, non-nucleoside inhibitors of HCV, and other
drugs for treating HCV, and combinations thereof.
More specifically, one or more compounds of the present invention
may be combined with one or more compounds selected from the group
consisting of 1) HIV protease inhibitors, e.g., amprenavir,
atazanavir, fosamprenavir, indinavir, lopinavir, ritonavir,
lopinavir+ritonavir, nelfinavir, saquinavir, tipranavir,
brecanavir, darunavir, TMC-126, TMC-114, mozenavir (DMP-450),
JE-2147 (AG1776), AG1859, DG35, L-756423, R00334649, KNI-272,
DPC-681, DPC-684, and GW640385X, DG17, PPL-100, 2) a HIV
non-nucleoside inhibitor of reverse transcriptase, e.g.,
capravirine, emivirine, delaviridine, efavirenz, nevirapine, (+)
calanolide A, etravirine, GW5634, DPC-083, DPC-961, DPC-963,
Mb-150, and TMC-120, TMC-278 (rilpivirine), efavirenz, BILR 355 BS,
VRX 840773, UK-453,061, RDEA806, 3) a HIV nucleoside inhibitor of
reverse transcriptase, e.g., zidovudine, emtricitabine, didanosine,
stavudine, zalcitabine, lamivudine, abacavir, amdoxovir,
elvucitabine, alovudine, Mb-210, racivir (.+-.-FTC), D-d4FC,
emtricitabine, phosphazide, fozivudine tidoxil, fosalvudine
tidoxil, apricitibine (AVX754), amdoxovir, KP-1461,
abacavir+lamivudine, abacavir+lamivudine+zidovudine,
zidovudine+lamivudine, 4) a HIV nucleotide inhibitor of reverse
transcriptase, e.g., tenofovir, tenofovir disoproxil
fumarate+emtricitabine, tenofovir disoproxil
fumarate+emtricitabine+efavirenz, and adefovir, 5) a HIV integrase
inhibitor, e.g., curcumin, derivatives of curcumin, chicoric acid,
derivatives of chicoric acid, 3,5-dicaffeoylquinic acid,
derivatives of 3,5-dicaffeoylquinic acid, aurintricarboxylic acid,
derivatives of aurintricarboxylic acid, caffeic acid phenethyl
ester, derivatives of caffeic acid phenethyl ester, tyrphostin,
derivatives of tyrphostin, quercetin, derivatives of quercetin,
S-1360, zintevir (AR-177), L-870812, and L-870810, MK-0518
(raltegravir), BMS-707035, MK-2048, BA-011, BMS-538158, GSK364735C,
6) a gp41 inhibitor, e.g., enfuvirtide, sifuvirtide, FB006M,
TRI-1144, SPC3, DES6, Locus gp41, CovX, and REP 9, 7) a CXCR4
inhibitor, e.g., AMD-070, 8) an entry inhibitor, e.g., SPO1A,
TNX-355, 9) a gp120 inhibitor, e.g., BMS-488043 and BlockAide/CR,
10) a G6PD and NADH-oxidase inhibitor, e.g., immunitin, 10) a CCR5
inhibitor, e.g., aplaviroc, vicriviroc, INCB9471, PRO-140,
INCB15050, PF-232798, CCR5mAb004, and maraviroc, 11) an interferon,
e.g., pegylated rIFN-alpha 2b, pegylated rIFN-alpha 2a, rIFN-alpha
2b, IFN alpha-2b XL, rIFN-alpha 2a, consensus IFN alpha, infergen,
rebif, locteron, AVI-005, PEG-infergen, pegylated IFN-beta, oral
interferon alpha, feron, reaferon, intermax alpha, r-IFN-beta,
infergen+actimmune, IFN-omega with DUROS, and albuferon, 12)
ribavirin analogs, e.g., rebetol, copegus, VX-497, and viramidine
(taribavirin) 13) NS5a inhibitors, e.g., A-831, A-689 and
BMS-790052, 14) NS5b polymerase inhibitors, e.g., NM-283,
valopicitabine, R1626, PSI-6130 (R1656), IDX184, PSI-7851, HCc-796,
BILB 1941, MK-0608, NM-107, R7128, VCH-759, PF-868554, GSK625433,
and XTL-2125, 15) NS3 protease inhibitors, e.g., SCH-503034
(SCH-7), VX-950 (Telaprevir), ITMN-191, and BILN-2065, 16)
alpha-glucosidase 1 inhibitors, e.g., MX-3253 (celgosivir) and
UT-231B, 17) hepatoprotectants, e.g., IDN-6556, ME 3738, MitoQ, and
LB-84451, 18) non-nucleoside inhibitors of HCV, e.g., benzimidazole
derivatives, benzo-1,2,4-thiadiazine derivatives, and phenylalanine
derivatives, 19) other drugs for treating HCV, e.g., zadaxin,
nitazoxanide (alinea), BIVN-401 (virostat), DEBIO-025, VGX-410C,
EMZ-702, AVI 4065, bavituximab, oglufanide, PYN-17, KPE02003002,
actilon (CPG-10101), KRN-7000, civacir, GI-5005, ANA-975, XTL-6865,
ANA 971, NOc-205, tarvacin, EHC-18, and NIM811, 19) pharmacokinetic
enhancers, e.g., BAS-100, SPI-452, PF-4194477, TMC-41629, GS-9350,
GS-9585, and roxythromycin, 20) RNAse H inhibitors, e.g., ODN-93
and ODN-112, 21) other anti-HIV agents, e.g., VGc-1, PA-457
(bevirimat), ampligen, HRG214, cytolin, polymun, VGX-410, KD247,
AMZ 0026, CYT 99007, A-221 HIV, BAY 50-4798, MDX010 (iplimumab),
PBS119, ALG889, and PA-1050040.
In a specific aspect of this embodiment, the additional therapeutic
agent is selected from ribavirin, telaprevir, boceprevir and
sofosbuvir (GS-7977 (formerly PSI-7977)).
A combination therapy described herein may be administered as a
simultaneous or sequential regimen. When administered sequentially,
the combination may be administered in two or more
administrations.
Co-administration of a compound of the invention with one or more
other active agents generally refers to simultaneous or sequential
administration of a compound of the invention and one or more other
active agents, such that therapeutically effective amounts of the
compound of the invention and one or more other active agents are
both present in the body of the patient.
Method for Treating Viral Infection
The present application provides a method for treating a
Flaviviridae viral infection comprising administering a
therapeutically effective amount of a compound described herein or
a pharmaceutically acceptable salt, isotope, stereoisomer, mixture
of stereoisomers, tautomer, ester or prodrug thereof, to a human
subject in need thereof.
Also provided is a method for treating a Coronaviridae viral
infection comprising administering a therapeutically effective
amount of a compound described herein or a pharmaceutically
acceptable salt, isotope, stereoisomer, mixture of stereoisomers,
tautomer, ester or prodrug thereof, to a human subject in need
thereof.
In one embodiment, the method of inhibiting or treating a disease
comprises administering to an animal a composition comprising an
effective amount of one or more compounds of the invention or a
pharmaceutically acceptable salt, isotope, stereoisomer, mixture of
stereoisomers, tautomer, ester or prodrug thereof and a
pharmaceutically acceptable carrier. The composition to be
administered may further contain a secondary therapeutic agent as
described above.
A method of the present application is particularly suitable for
use with humans, but may be used with other animals, particularly
mammals, such as, for example, non-human primates, companion
animals, farm animals, laboratory animals, and wild and zoo
animals.
A method of the present application is particularly useful to treat
diseases caused directly or indirectly by Flaviviridae virus since
the compounds of the present invention have inhibitory activity
against those viruses. In some embodiments, therefore, a method of
the present invention is used in inhibiting or treating diseases
caused by a Hepatitis C virus. In some embodiments, therefore, a
method of the present invention is used in inhibiting or treating
diseases caused by a Hepatitis B virus. In an aspect, such a method
is applied to a patient with a disease caused by the viral
infection such as dengue fever, yellow fever, hepatitis C, Japanese
encephalitis, Kyasanur forest disease, Murray valley encephalitis,
St. Louis encephalitis, tick-borne encephalitis or West Nile
encephalitis.
In some embodiments, a sustained virologic response is achieved at
about 12 weeks, at about 10 weeks, at about 8 weeks, at about 6
weeks, or at about 4 weeks, or at about 4 months, or at about 5
months, or at about 6 months, or at about 1 year, or at about 2
years.
A method of the present application is also particularly useful to
treat diseases caused directly or indirectly by Coronaviridae virus
since the compounds of the present invention have inhibitory
activity against those viruses. In some embodiments, therefore, a
method of the present invention is used in inhibiting or treating
diseases caused by a SARS coronarirus. In an aspect, such a method
is applied to a patient with a disease caused by the viral
infection such as severe acute respiratory syndrome (SARS), cancer,
inflammation, obesity, acquired immune deficiency syndrome (AIDS),
or cirrhosis.
In another aspect, the compounds disclosed herein can be used for
treating cancer. In yet another aspect, the compounds disclosed
herein can be used for immunomodulation. In some embodiments,
therefore, a method of the present invention comprises adjusting an
immune response to a desired level, as in immunopotentiation,
immunosuppression, or induction of immunologic tolerance.
In some embodiments, the compound is administered for about 12
weeks. In further embodiments, the compound is administered for
about 12 weeks or less, for about 10 weeks or less, for about 8
weeks or less, for about 6 weeks or less, or for about 4 weeks or
less. The compound may be administered once daily, twice daily,
once every other day, two times a week, three times a week, four
times a week, or five times a week.
In another aspect, the compounds disclosed herein can be used for
treating cancer. In yet another aspect, the compounds disclosed
herein can be used for immunomodulation. In some embodiments,
therefore, a method of the present invention comprises adjusting an
immune response to a desired level, as in immunopotentiation,
immunosuppression, or induction of immunologic tolerance.
EXAMPLES
The following examples are merely illustrative, and do not limit
this disclosure in any way.
List of Abbreviations and Acronyms
TABLE-US-00006 Abbreviation Meaning .degree. C. Degree Celsius Ac
Acetyl app Apparent Aq Aqueous Boc tert-Butoxycarbonyl Br Broad Bu
Butyl cat Catalytic CBS Corey Bakshi Shibata CDMT
2-chloro-4,6-dimethoxy-1,3,5-triazine cm Centimeter COMU
(1-Cyano-2-ethoxy-2- oxoethylidenaminooxy)dimethylamino-
morpholino-carbenium hexafluorophosphate CP/Cp Cyclopentyl CPME
Cyclopentyl methyl ether Cy/cHex Cyclohexyl d Doublet DBU
1,8-Diazabicyclo[5.4.0]undec-7-ene DCC
N,N'-dicyclohexylcarbodiimide DCE Dichloroethane DCM
Dichloromethane dd Doublet of doublets ddd Doublet of doublet of
doublet ddt Doublet of doublet of triplet DIAD Diisopropyl
azodicarboxylate DIPEA N,N-Diisopropylethylamine DMAP
4-Dimethylaminopyridine DME 1,2-dimethoxyethane DMF
Dimethylformamide DMSO Dimethylsulfoxide dppf
1,1'-bis(diphenylphosphino)ferrocene dq Doublet of quartet dt
Doublet of triplet dtd Doublet of triplet of doublet EDC
1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Equiv/eq Equivalents
Et Ethyl g Grams gen. Generation HATU
(Dimethylamino)-N,N-dimethyl(3H-[1,2,3]triazolo[4,5-
b]pyridin-3-yloxy)methaniminium hexafluorophosphate HBTU
2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate HMDS Hexamethyldisilazane HMPA
hexamethylphosphoramide HOBT Hydroxybenzotriazole HPLC
High-performance liquid chromatography hrs/h Hours Hz Hertz
IC.sub.50 The half maximal inhibitory concentration Im imidazole
i-Pr/iPr Isopropyl J Coupling constant Kg Kilogram LCMS Liquid
chromatography-mass spectrometry LDA Lithium diisopropylamide M
Molar m Multiplet m/z mass-to-charge ratio M+ Mass peak Me Methyl
mg Milligram MHz Megahertz min Minute mL Milliliter mM Millimolar
mm Millimeter mmol Millimole mol Mole Ms Methanesulfonyl MW
Microwave N normal nM Nanomolar NMM N-methylmorpholine NMR Nuclear
magnetic resonance nPrOH n-propanol o-Tol o-Tolyl Ph Phenyl pTSA
p-Toluenesulfonic acid Py/pyr Pyridine PyAOP
7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium
hexafluorophosphate PyBop
benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate
q Quartet Rf Retention factor RT/rt/r.t. Room temperature s Singlet
sat. Saturated t Triplet TBAF Tetra-n-butylammonium fluoride
TBDMS/TBS tert-Butyldimethylsilyl TBDPS tert-Butyldiphenylsilyl
t-Bu tert-butyl td Triplet of doublets TEA Triethylamine Tf
Trifluoromethanesulfonyl TFA Trifluoroacetic acid THF
Tetrahydrofuran TLC Thin layer chromatography TMEDA
tetramethylethylenediamine TMS Trimethylsilyl Tr/tr Retention time
Ts Tosyl tt Triplet of triplet UV Ultraviolet wt. weight .delta.
Chemical shift .mu.L Microliter .mu.M Micromolar .mu.mol
Micromole
Example 1:
(E)-(2R,5S,11S,14S,17R,18R)-18-Hydroxy-14-isopropyl-2,11,17-tri-
methyl-3,9,12,15,28-pentaaza-tricyclo[21.3.1.1*5,9]octacosa-1(26),21,23(27-
),24-tetraene-4,10,13,16-tetraone
##STR00042##
A solution of
1-((1R,5S)-10,10-dimethyl-3,3-dioxo-3lambda*6*-thia-4-aza-tricyclo[5.2.1.-
0*1,5*]dec-4-yl)-propan-1-one (3.95 g, 14.55 mmol) in toluene (50
mL) was prepared, then evaporated to dryness. This process was
repeated and the resulting white solid was dissolved in anhydrous
dichloromethane (16 mL). A small quantity of calcium hydride was
added before adding tert-butyldimethylsilyl
trifluoromethanesulfonate (3.83 mL, 14.5 mmol) and anhydrous
triethylamine (2.33 mL, 16.7 mmol). The reaction mixture was
stirred at RT ("RT") under a nitrogen atmosphere for 15 hours
("h"). The resulting solution was evaporated to yield a thick
paste, which was re-dissolved in anhydrous dichloromethane (15 mL)
and added dropwise to a stirred solution of 4-pentenal (2.69 g,
32.0 mmol) and titanium tetrachloride (1 M in dichloromethane, 32
mL, 32 mmol) in anhydrous dichloromethane (20 mL) at -78.degree.
C., under a nitrogen atmosphere. The reaction was stirred at
-78.degree. C. for 30 minutes ("min") before diluting with
saturated aqueous ammonium chloride solution (100 mL). The layers
were separated and the aqueous layer was extracted with
dichloromethane (2.times.50 mL). The combined extract was dried
over sodium sulfate, filtered and evaporated to give a brown gum.
This was purified by silica gel chromatography using
iso-hexanes/ethyl acetate 4:1 to yield the title compound (3.09 g,
60%) as a colorless gum.
##STR00043##
A solution of 1a (12.0 g, 0.034 mol) in anhydrous dichloromethane
(520 mL) was cooled to 0.degree. C., before adding pyridine (5.5
mL, 0.068 mol) then tert-butyldimethylsilyl
trifluoromethanesulfonate (9 mL, 0.039 mol). The reaction mixture
was stirred at 0.degree. C. for 15 min then allowed to warm to RT
and stirred for a further 1.5 h. The reaction mixture was washed
with saturated sodium bicarbonate (400 mL). The aqueous wash was
back-extracted with dichloromethane (200 mL). The organic layers
were combined and washed with dilute brine (200 mL) and 2 M
hydrochloric acid (200 mL). The solution was dried over sodium
sulfate, filtered and evaporated to give the title product (15.29
g, 96%) as a white solid.
##STR00044##
A solution of 1b (15.29 g, 0.0325 mol) in tetrahydrofuran (300 mL)
was prepared and a 2 M aqueous solution of lithium hydroxide (120
mL) was added. The stirred mixture was heated to 60.degree. C. for
15 h. The reaction was diluted with 2 M hydrochloric acid (250 mL).
The layers were separated and the aqueous was extracted with ethyl
acetate (2.times.200 mL). The organic layers were combined, dried
over sodium sulfate, filtered and evaporated to give a cream solid
(16.7 g). The solid was purified by silica gel chromatography using
3:7 ethyl acetate/iso-hexanes to yield the title product (7.18 g,
81%) as a colorless gum.
##STR00045##
A solution of (S)-2-tert-butoxycarbonylamino-propionic acid (3.28
g, 17.32 mmol) in acetonitrile (160 mL) was cooled to 0.degree. C.
before addition of N,N-diisopropylethylamine (12 mL, 69.3 mmol)
then 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate (6.57 g, 17.32 mmol). The reaction mixture was
stirred at 0.degree. C. for 20 min and a solution of
(S)-hexahydropyridazine-3-carboxylic acid 2,2,2-trichloroethyl
ester trifluoroacetic acid salt (preparation described in Angew.
Chem. Int. Ed. Engl. 1999, 38, 2443, 6.49 g, 17.3 mmol) in
acetonitrile (80 mL) was added. The reaction was allowed to warm to
RT and was stirred for 15 h. The reaction mixture was evaporated
then re-dissolved in ethyl acetate (150 mL). The solution was
washed with brine (150 mL). The brine was back extracted with ethyl
acetate (50 mL). The organic layers were combined, dried over
sodium sulfate, filtered and evaporated to give a dark oil. The oil
was purified by silica gel chromatography using iso-hexanes/ethyl
acetate 1:1 to yield the title compound (6.88 g, 92%) as a
colorless gum.
##STR00046##
A solution of 1d (6.88 g, 15.9 mmol) in dichloromethane (200 mL)
was prepared and trifluoroacetic acid (50 mL) was added. The
reaction mixture was stirred at RT for 2 h. TLC showed the reaction
to be complete. The solution was evaporated to give a brown oil.
This was azeotroped with toluene (50 mL) and the resultant oil was
dried under vacuum to give
(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic
acid 2,2,2-trichloro-ethyl ester trifluoroacetic acid salt (7.8 g)
as a brown gum. A solution of
((S)-1-carbamoyl-2-methylpropyl)-carbamic acid tert-butyl ester in
acetonitrile (300 mL) was cooled to 0.degree. C. before adding
N,N-diisopropylethylamine (13.8 mL, 79.7 mmol) and
2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate (6.33 g, 16.7 mmol). The reaction was stirred
at 0.degree. C. for 15 min before adding a solution of the
(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic
acid 2,2,2-trichloro-ethyl ester trifluoroacetic acid salt (ca.
15.9 mmol) in acetonitrile (85 mL). The reaction was stirred at
0.degree. C. for a further 20 min then allowed to warm to RT and
stirred for 15 h. The reaction mixture was evaporated then
re-dissolved in ethyl acetate (250 mL). The solution was washed
with water (150 mL) then dried over sodium sulfate, filtered and
evaporated to give a red oil. This was purified by silica gel
chromatography using iso-hexanes/ethyl acetate 7:3 then
iso-hexanes/ethyl acetate 1:1 to yield the title compound (8.2 g,
92%) as a pale orange amorphous solid.
##STR00047##
A solution of 1e (10.0 g, 18.5 mmol) in anhydrous dichloromethane
(200 mL) was prepared and trimethylsilyl trifluoromethanesulfonate
(5 mL, 27.75 mmol) was added. The reaction mixture was stirred at
RT for 2 h, then N,N-diisopropylethylamine (13.2 mL, 75.8 mmol) was
added and the reaction mixture was evaporated to dryness. The
residue was re-dissolved in acetonitrile (200 mL) and a solution of
(2R,3R)-3-(tert-butyl-dimethyl-silanyloxy)-2-methyl-hept-6-enoic
acid (5.04 g, 18.5 mmol) in acetonitrile (60 mL) was added followed
by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (5.0 g, 26.4 mmol)
and 1-hydroxybenzotriazole (4.4 g, 26.4 mmol). The reaction mixture
was stirred at RT for 15 h. It was evaporated to give a thick
yellow oil. The oil was purified by silica gel chromatography using
1:1 ethyl acetate/iso-hexanes then 3:2 ethyl acetate/iso-hexanes to
yield the title product (8.75 g, 69%) as a white solid.
##STR00048##
A solution of (R)-bromo-.alpha.-methylbenzylamine (1.023 g, 5.112
mmol) in dichloromethane (20 mL) was subsequently treated with
triethylamine (720 .mu.L, 5.112 mmol) and di-tert-butyl dicarbonate
(1.784 g, 8.179 mmol). After overnight stirring at RT, the
volatiles were removed in vacuo and the residue was purified by
silica gel chromatography using a 50 g Isolute cartridge eluted
with a continuous gradient of iso-hexanes/diethyl ether 1:0 to 4:1
to afford the title compound (1.552 g, 100%) as a white solid.
##STR00049##
A solution of 1g (10.26 g, 0.0342 mol.) and tributyl(vinyl)tin
(32.5 g, 30 mL, 0.103 mol.) in toluene (175 mL) was purged with
nitrogen for 30 min before addition of bis(triphenylphosphine)
palladium (II) dichloride (2.38 g, 0.0034 mol.). The stirred
mixture was heated to 60.degree. C. for 16 h before cooling to RT.
The reaction mixture was filtered through hyflo-supercel then
evaporated to give a dark coloured oil. The oil was purified by
silica gel chromatography using iso-hexanes/ethyl acetate 19:1 to
yield the title compound (6.95 g, 82%) as a yellow oil.
##STR00050##
A solution of 1h (6.95 g, 28.1 mmol.) in 1,4-dioxane (30 mL) was
prepared and a solution of hydrogen chloride in 1,4 dioxane (4 M,
60 mL) was added. The reaction mixture was stirred at RT for 2 h
then evaporated to dryness. The resultant solid was re-dissolved in
toluene and evaporated. The solid was triturated with diethyl
ether, which was removed by decanting. The solid was then dried
under vacuum to give the title compound (4.96 g, 96%) as an
off-white solid.
##STR00051##
A solution of 1f (8.75 g, 12.75 mmol) in tetrahydrofuran (350 mL)
was prepared and zinc powder (8.44 g, 127.5 mmol) was added
followed by a 1 M aqueous solution of ammonium acetate (90 mL, 90
mmol). The reaction mixture was vigorously stirred for 16 h then
filtered through hyflo-supercel. The solution was cooled over an
ice bath before addition of aqueous ammonium chloride solution (350
mL). It was allowed to re-cool before acidifying to pH 1 by
addition of 2 M hydrochloric acid. The layers were separated and
the aqueous was extracted with ethyl acetate (2.times.250 mL). The
organic layers were combined, dried over sodium sulfate filtered
and evaporated to give a white solid. The solid was azeotroped with
toluene (3.times.200 mL) then dried under vacuum to give a white
solid (6.16 g), which was dissolved in acetonitrile (400 mL) before
adding (R)-1-(3-vinyl-phenyl)-ethylamine hydrochloride (2.34 g,
12.75 mmol) followed by N,N-diisopropylethylamine (8.9 mL, 51 mmol)
and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (6.8 g, 17.85 mmol). The reaction
mixture was stirred at RT for 16 h and then evaporated to give a
brown gum. The gum was purified by silica gel chromatography using
ethyl acetate then 1:4 acetone/ethyl acetate to yield the title
compound (5.51 g, 76%) as a cream solid.
##STR00052##
A solution of 1j (0.50 g, 0.88 mmol) in anhydrous dichloromethane
(20 mL) was cooled over an ice bath, before adding pyridine (92
.mu.L, 1.14 mmol) and tert-butyldimethylsilyl
trifluoromethanesulfonate (242 .mu.L, 1.05 mmol). The reaction
mixture was stirred at 0.degree. C. for 15 min, then allowed to
warm to RT and stirred for 1 hour. The reaction mixture was
evaporated and the residue was purified by silica gel
chromatography using ethyl acetate to yield the title product (477
mg, 80%) as a white solid.
##STR00053##
A solution of 1k (477 mg, 0.7 mmol) in 1,2-dichloroethane (250 mL)
was prepared and Hoveyda-Grubbs 2.sup.nd generation catalyst (43
mg, 0.07 mmol) was added. The stirred reaction mixture was heated
to 80.degree. C. for 1 hour. The reaction mixture was cooled to RT
before adding silica gel. The mixture was stirred for 10 min then
evaporated and the residue was purified by silica gel
chromatography using ethyl acetate to yield the title product (198
mg, 43%) as a white solid.
##STR00054##
A solution of 1l (198 mg, 0.3 mmol) in tetrahydrofuran (20 mL) was
cooled over an ice bath before adding a 1 M solution of
tetrabutylammonium fluoride in tetrahydrofuran (1.5 mL, 1.5 mmol).
The reaction was allowed to warm to RT and was stirred for 1 hour.
The reaction mixture was treated with saturated aqueous sodium
bicarbonate solution (20 mL) and extracted with ethyl acetate
(2.times.20 mL). The extract was dried over sodium sulfate,
filtered and evaporated. The residue was purified by silica gel
chromatography using 1:9 acetone/ethyl acetate to yield the title
product (150 mg, 92%) as a white solid. .sup.1H NMR (300 MHz,
d.sub.6-DMSO) .delta. 0.83 (d, J=6.5 Hz, 3H), 0.85 (d, J=6.5 Hz,
3H), 1.12-1.21 (m, 4H), 1.28 (d, J=7.1 Hz, 3H), 1.36 (d, J=6.9 Hz,
3H), 1.40-1.90 (m, 7H), 1.92-2.08 (m, 1H), 2.19-2.34 (m, 2H),
2.67-2.80 (m, 1H), 3.56-3.65 (m, 1H), 3.99-4.12 (m, 1H), 4.22 (br
d, J=12.2 Hz, 1H), 4.75 (d, J=11.8 Hz, 1H), 4.87-4.99 (m, 1H),
5.12-5.24 (m, 1H), 5.40 (d, J=4.5 Hz, 1H), 6.14-6.33 (m, 2H),
7.10-7.35 (m, 5H), 7.88 (d, J=8.3 Hz, 1H), 8.55 (d, J=8.3 Hz, 1H).
LCMS (m/z) 542.3 [M+H], Tr=1.87 min.
Example 2:
(E)-(2R,5S,11S,14S,17R,18R)-14-Isopropyl-2,11,17-trimethyl-18-(-
2,2,2-trifluoro-ethoxy)-3,9,12,15,28-pentaaza-tricyclo[21.3.1.1*5,9]octaco-
sa-1(26),21,23(27),24-tetraene-4,10,13,16-tetraone
##STR00055##
A solution of hexamethyldisilazane (235 .mu.L, 1.13 mmol) in
anhydrous tetrahydrofuran (4 mL) was cooled to -10.degree. C.
before adding a 2.5 M solution of n-butyllithium (380 .mu.L, 0.94
mmol). The stirred mixture was warmed to 0.degree. C. for 10 min
before cooling to -78.degree. C. A solution of Compound 1 (102 mg,
0.188 mmol) in anhydrous dimethylformamide (1.5 mL) and anhydrous
tetrahydrofuran (1.5 mL) was added dropwise over 2 min followed by
the addition of 2,2,2-trifluoroethyl trifluoromethanesulfonate (135
.mu.L, 0.94 mmol). The reaction mixture was stirred at -78.degree.
C. under a nitrogen atmosphere then allowed to gradually warm to
RT. The reaction mixture was quenched with the addition of a
saturated aqueous solution of ammonium chloride (15 mL). The
mixture was extracted with ethyl acetate (2.times.15 mL). The
extract was dried over sodium sulfate, filtered and evaporated. The
residue was purified by silica gel chromatography using 1:9
acetone/ethyl acetate to give a colorless gum (65 mg). The gum was
further purified by reverse phase chromatography, using a 10 g C18
cartridge eluted with 2:3 acetonitrile/water. The partially
evaporated fractions were extracted with ethyl acetate (2.times.15
mL). The extract was dried over sodium sulfate, filtered and
evaporated to yield the title product (17 mg, 30%) as a white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3) 0.92 (d, J=6.7 Hz, 3H),
0.96 (d, J=6.9 Hz, 3H), 1.32 (d, J=7.1 Hz, 3H), 1.39 (d, J=7.1 Hz,
3H), 1.50 (d, J=6.7 Hz, 3H), 1.74-1.85 (m, 1H), 1.86-2.08 (m, 3H),
2.10-2.40 (m, 3H), 2.57-2.78 (m, 2H), 3.19-3.31 (m, 1H), 3.49 (app
t, J=6.7 Hz, 1H), 3.58-3.79 (m, 4H), 3.81-4.18 (m, 2H), 4.49 (br d,
J=12.3 Hz, 1H), 5.03-5.17 (m, 1H), 5.32-5.47 (m, 1H), 6.14-6.30 (m,
1H), 6.37-6.72 (m, 4H), 7.06-7.30 (m, 4H). LCMS (m/z) 624.3 [M+H],
Tr=2.57 min.
Example 3:
(2R,5S,11S,14S,17R,18R)-18-Hydroxy-14-isopropyl-2,11,17-trimeth-
yl-3,9,12,15,28-pentaaza-tricyclo[21.3.1.1*5,9]octacosa-1(26),23(27),24-tr-
iene-4,10,13,16-tetraone
##STR00056##
A solution of Compound 1 (100 mg, 0.184 mmol) in methanol (5 mL)
was prepared and 10% palladium on carbon (5 mg) was added. The
stirred mixture was placed under a hydrogen atmosphere for 1.5 h.
The reaction mixture was filtered through hyflo-supercel then
through a 0.2 .mu.m filter before evaporating to give the title
compound (95 mg, 95%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) 0.93 (d, J=7.1 Hz, 3H), 0.96 (d, J=7.1 Hz, 3H), 1.32
(d, J=6.7 Hz, 3H), 1.37-2.16 (m, 12H), 1.54 (d, J=6.9 Hz, 3H),
2.34-2.46 (m, 1H), 2.52-2.75 (m, 3H), 3.16-3.27 (m, 1H), 3.43-3.62
(m, 3H), 3.74 (d, J=11.8 Hz, 1H), 3.98-4.06 (m, 1H), 4.50 (d,
J=13.2 Hz, 1H), 5.11-5.36 (m, 2H), 6.39-6.50 (m, 2H), 7.70 (d,
J=8.5 Hz, 1H), 7.06-7.28 (m, 4H). LCMS (m/z) 544.3 [M+H], Tr=1.91
min.
Example 4
##STR00057##
To a suspension of sodium hydride (60% in mineral oil, 783 mg, 19.6
mmol) in dry tetrahydrofuran at 0.degree. C., under nitrogen was
added dropwise triethyl phosphonoacetate (3.38 mL, 19.6 mmol). The
white suspension was stirred for 1 h where it became a solution,
then 2,2-dimethyl-4-pentenal (2.42 mL, 17.8 mmol) was slowly added
and the resulting green/yellow solution was stirred at 0.degree. C.
and allowed to warm to RT. After 72 h ethanol (1 mL) was added to
the solution, followed by water (100 mL) and the organics were
extracted with diethyl ether (2.times.200 mL). The combined
organics were washed with water (200 mL), dried over magnesium
sulfate, filtered and concentrated in vacuo to give a crude
residue. This was purified by silica gel chromatography, using
iso-hexanes (144 mL), then iso-hexanes/diethyl ether 50:50 (72 mL),
then diethyl ether (48 mL) to give the title compound (3.20 g, 99%)
as an oil.
##STR00058##
To 4a (3.20 g, 17.5 mmol) in dry methanol at RT, under nitrogen was
slowly added magnesium powder (1.28 g, 52.7 mmol) while monitoring
the temperature rise. After the addition was complete the mixture
was stirred at RT overnight. After this time, to the reaction was
added additional magnesium powder (852 mg, 35.1 mmol) and the
reaction mixture stirred for 2 h. The mixture was neutralized from
pH 10 to pH 7 with 2 M hydrochloric acid and then concentrated in
vacuo to give a residue. The residue was suspended in ethyl acetate
(300 mL) and water (500 mL) and the layers separated. The aqueous
phase was then re-extracted with ethyl acetate (200 mL) and the
combined organics were dried over magnesium sulfate, filtered and
concentrated in vacuo to give the title compound (1.96 g, 61%) as a
colorless oil.
##STR00059##
To 4b (2.02 g, 11.0 mmol) in a mixture of tetrahydrofuran (35 mL),
methanol (9 mL) and water (9 mL) at RT, was added lithium hydroxide
monohydrate (1.38 g, 32.9 mmol) and the mixture was stirred at RT
for 1 h. After this time additional lithium hydroxide monohydrate
(460 mg, 11.0 mmol) was added and the reaction mixture stirred for
1 h. After this time more lithium hydroxide monohydrate (460 mg,
11.0 mmol) was added and the reaction mixture stirred for 30 min.
The reaction mixture was then concentrated in vacuo and the residue
diluted with water (200 mL) and the organics extracted with diethyl
ether (3.times.50 mL). The aqueous phase was acidified from pH 14
to pH 1 with 2 M hydrochloric acid and the organics extracted with
ethyl acetate (3.times.50 mL). The combined ethyl acetate organics
were then dried over sodium sulfate, filtered and concentrated in
vacuo to give the desired product (1.03 g, 60%) as an oil.
##STR00060##
A solution of 1e (1.06 g, 2.00 mmol) in dichloromethane (10 mL) was
cooled in an ice-water bath under nitrogen. Trimethylsilyl
trifluoromethanesulfonate (0.591 mL, 4.00 mmol) was added dropwise,
and the resulting solution was stirred for 1.5 h. The reaction was
quenched with N,N-diisopropylethylamine (1.4 mL, 8.00 mmol) and the
reaction mixture was concentrated in vacuo to give
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester a white solid
which was used without further purification. To
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester generated in
the previous step was added a solution of 4,4-dimethyl-hept-6-enoic
acid (343 mg, 2.20 mmol) in acetonitrile (70 mL). To this mixture
was added N,N-diisopropylethylamine (1.4 mL, 8.03 mmol) and the
reaction cooled in an ice-water bath before the addition of
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate methanaminium (1.07 g, 2.80 mmol). The yellow
solution was then stirred and allowed to warm to RT overnight. The
reaction mixture was quenched with 2 M hydrochloric acid (40 mL)
and concentration in vacuo. To the residue was added water (200 mL)
and the organics were extracted with ethyl acetate (3.times.150
mL). The combined organics were then washed with brine (100 mL) and
sodium hydrogen carbonate (200 mL). The organics were then dried
over magnesium sulfate, filtered and concentrated in vacuo to give
a crude yellow oil (2.27 g). This was purified by silica gel
chromatography using iso-hexanes/ethyl acetate 90:10 (300 mL) then
iso-hexanes/acetone 50:50 (300 mL) then iso-hexanes/acetone 0:100
to give the title compound (1.10 g, 97%) as a colorless oil.
##STR00061##
A solution of 4d (1.10 g, 1.94 mmol) in tetrahydrofuran (40 mL) was
prepared and zinc powder (2.79 g, 42.6 mmol) was added followed by
a solution of ammonium acetate (2.24 g, 29.0 mmol) in water (10
mL). The reaction mixture was stirred at RT for 24 h. Saturated
aqueous potassium hydrogen sulphate (pH 2, 30 mL) and ethyl acetate
(50 mL) were added and the suspension filtered through
hyflo-supercel washing through with ethyl acetate. The layers were
separated. And the aqueous phase was extracted with ethyl acetate
(3.times.100 mL). The combined organic layers were washed with
brine (250 mL), dried over sodium sulfate, filtered and evaporated
to give a colorless gum. The residue was azeotroped with toluene
(3.times.100 mL) to give the title compound (816 mg, 96%) as a
white solid.
##STR00062##
4e (439 mg, 1.00 mmol) was suspended in acetonitrile (35 mL) and
(R)-1-(3-vinyl-phenyl)-ethylamine hydrochloride (202 mg, 1.10 mmol)
was added followed by N,N-diisopropylethylamine (700 .mu.L, 5.00
mmol) and the mixture cooled in an ice-water bath before addition
of 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate methanaminium (532 mg, 1.40 mmol). The reaction
mixture was stirred and allowed to warm to RT over 72 h. To the
reaction mixture was added 2 M hydrochloric acid (20 mL) and the
mixture concentrated in vacuo to give a residue. The aqueous layer
was extracted with ethyl acetate (3.times.100 mL) and the combined
ethyl acetate layers were washed with brine followed by sodium
hydrogen carbonate (100 mL) then dried over sodium sulfate,
filtered and concentrated in vacuo to give a crude residue. The
residue was purified by silica gel chromatography using
iso-hexanes/ethyl acetate 50:50 then neat ethyl acetate to give the
title compound (402 mg, 71%).
##STR00063##
To a solution of 4f (402 mg, 0.71 mmol) in 1,2-dichloroethane (210
mL) was added Hoveyda-Grubbs 2.sup.nd generation catalyst (45 mg,
0.07 mmol) and the reaction mixture heated at 80.degree. C. for 1
h. After this time the mixture was allowed to cool to RT and
concentrated in vacuo. The residue was purified by silica gel
chromatography using a gradient of ethyl acetate to ethyl
acetate/acetone 75:25. Impure product (240 mg) was collected which
was further purified by silica gel chromatography using ethyl
acetate. After repurification, product (43 mg) was collected
containing minor impurities. The impurities were removed via
trituration from diethyl ether to give the title compound (30 mg,
8%) as a white solid. .sup.1H NMR (300 MHz, CD.sub.3OD) 0.89 (d,
J=6.7 Hz, 3H), 0.92 (d, J=6.7 Hz, 3H), 0.96 (s, 3H), 0.97 (s, 3H),
1.35 (d, J=6.9 Hz, 3H), 1.50 (d, J=7.1 Hz, 3H), 1.59-1.83 (m, 3H),
1.85-2.00 (m, 3H), 2.09 (d, J=7.1 Hz, 2H), 2.15-2.25 (m, 1H),
2.31-2.44 (m, 1H), 2.78 (td, J=2.7, 12.7 Hz, 1H), 3.40-3.72 (m,
2H), 4.05-4.17 (m, 1H), 4.34 (brd, J=13.2 Hz, 1H), 5.05 (q, J=7.1
Hz, 1H), 5.35 (q, J=7.1 Hz, 1H), 6.19-6.32 (m, 1H), 6.37 (d, J=15.8
Hz, 1H), 7.15 (d, J=7.6 Hz, 1H), 7.21-7.29 (m, 2H), 7.37 (d, J=7.8
Hz, 1H). LCMS (m/z) 540.3 [M+H], Tr=2.41 min.
Example 5
##STR00064##
A solution of 6-bromo-3-chloro-isoquinoline (8.0 g, 33 mmol) and
tributyl-(1-ethoxyvinyl)-tin (14.88 g, 14 mL, 41.2 mmol) in toluene
(100 mL) was degassed with nitrogen for 30 min.
Bis(triphenylphosphine)palladium(II) dichloride (1.16 g, 1.65 mmol,
5 mol %) was added and the reaction mixture was heated at
60.degree. C. for 20 h. The reaction mixture was cooled to RT, the
mixture was filtered and the filtrate was evaporated. The residue
was purified by silica gel chromatography using a gradient of
iso-hexanes/ethyl acetate 20:1 to 10:1 to afford the title compound
(7.1 g, 92%) as a yellow solid.
##STR00065##
A solution of 5a (7.1 g, 30 mmol) in 1,4-dioxane (60 mL) and 2 M
hydrochloric acid (30 mL) was stirred at RT for 30 min. The
majority of the solvent was evaporated and the residue was
partitioned between ethyl acetate and water. The organic extracts
were combined, washed with water and brine, dried over sodium
sulfate, filtered and evaporated. The residue was triturated with
5% ether in iso-hexanes and the resulting solid was collected and
dried to afford the title compound (6.0 g, 97%) as a white
solid.
##STR00066##
A solution of 5b (1.72 g, 8.3 mmol) in tetrahydrofuran (40 mL) was
stirred under nitrogen. Titanium (IV) ethoxide (3.8 g, 3.45 mL,
16.6 mmol, tech. grade) was added followed by
(R)-(+)-2-methyl-propanesulfinimide (1.11 g, 9.2 mmol) and the
reaction mixture was stirred at 60.degree. C. under nitrogen for 18
h. Additional (R)-(+)-2-methyl-propanesulfinimide (190 mg, 1.67
mmol) was added and the reaction mixture was stirred at 65.degree.
C. for further 2 h. The reaction mixture was cooled to RT and ethyl
acetate and brine were added. The suspension was filtered through
Celite and the filter pad was washed with ethyl acetate. The ethyl
acetate layer was separated, washed with brine, dried over sodium
sulfate, filtered and evaporated. The residue was purified by
silica gel chromatography using a gradient of iso-hexanes/ethyl
acetate 7:3 to 3:7 to afford the title compound (2.2 g, 86%) as a
yellow solid.
##STR00067##
A mixture of (1S,2R)-(-)-cis-1-amino-2-indanol (60 mg, 0.4 mmol),
dichloro(p-cymene)ruthenium(II) dimer (122 mg, 0.2 mmol) and
powdered 4 {acute over (.ANG.)} molecular sieves (2 g) was
suspended in anhydrous 2-propanol (9 mL) and stirred under
nitrogen. The suspension was heated at 90.degree. C. for 20 min.
The reaction mixture was cooled to 40.degree. C. and a solution of
5c (1.23 g, 4 mmol) in 2-propanol (28 mL) was added followed by a
solution of potassium tert-butoxide (122 mg, 1.1 mmol) in
2-propanol (10 mL). The reaction mixture was stirred for 2 h at
40.degree. C. and then allowed to cool. The mixture was poured
directly onto a silica gel cartridge and eluted with ethyl acetate
to give, after evaporation, the title compound (1.19 g, 96%) as a
brown gum.
##STR00068##
A mixture of 5d (0.66 g, 2.11 mmol), tributyl(vinyl)tin (1.85 mL,
6.35 mmol) and palladium tetrakis(triphenylphosphine) (488 mg, 0.42
mmol) in 1,4-dioxane (10.5 mL) was capped in a microwave vial. The
reaction mixture was irradiated and stirred at 160.degree. C. for
40 min in a microwave reactor. A second reaction was carried under
identical scale and conditions and the reaction mixtures combined
and evaporated. The residue was purified by silica gel
chromatography using a gradient of 1:3 to 1:0 ethyl
acetate/iso-hexanes to afford the title compound (1 g) as a brown
gum. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.1.27 (s, 9H), 1.62
(d, J=7.7 Hz, 3H), 3.53 (br s, 1H), 4.69-4.77 (m, 1H), 5.51 (dd,
J=10.7, 1.3 Hz, 1H), 6.39 (dd, J=17.4, 1.3 Hz, 1H), 6.95 (dd,
J=17.2, 10.7 Hz, 1H), 7.55-7.58 (m, 2H), 7.75 (s, 1H), 7.95 (d,
J=8.3 Hz, 1H), 9.20 (s, 1H). LCMS (m/z) 303.0 [M+H], Tr=1.48
min.
##STR00069##
5e was suspended in 4 M hydrochloric acid in 1,4-dioxane (17 mL, 68
mmol) and methanol was added (34 mL). The reaction mixture was
stirred for 90 min and then evaporated. The residue was passed
through a SCX cartridge eluting with methanol and then methanolic
ammonia. The basic fraction was collected and evaporated to give
the title compound (530 mg, 63% over 2 steps) as a beige solid.
##STR00070##
A solution of
1-((1R,5S)-10,10-dimethyl-3,3-dioxo-3lambda*6*-thia-4-aza-tricyclo[5.2.1.-
0*1,5*]dec-4-yl)-propan-1-one (6.0 g, 22.1 mmol) in anhydrous
dichloromethane (24 mL) was prepared and tert-butyldimethylsilyl
trifluoromethanesulfonate (5.0 mL, 22.1 mmol) was added, followed
by anhydrous triethylamine (3.54 mL, 25.4 mmol). The reaction
mixture was stirred at RT under a nitrogen atmosphere for 15 h.
This gave a dark solution that was evaporated to give an oil. The
oil was dissolved in anhydrous dichloromethane (22 mL) and the
solution was added dropwise to a solution of crotonaldehyde (3.66
mL, 44.2 mmol) and titanium tetrachloride (1 M in dichloromethane,
44.2 mL, 44.2 mmol) in dichloromethane (22 mL) at -78.degree. C.
under a nitrogen atmosphere. The reaction mixture was stirred at
-78.degree. C. for 1 hour, before addition of ammonium chloride
solution (30 mL). The stirred mixture was allowed to warm to RT
before separating the layers. The aqueous layer was extracted with
dichloromethane (2.times.25 mL). The organic layers were combined,
dried over sodium sulfate, filtered and evaporated to give a brown
oil. The oil was purified by silica gel chromatography using
iso-hexanes/ethyl acetate 4:1 to yield the title compound (6.7 g,
89%) as a colorless solid.
##STR00071##
A solution of 5g (4.15 g, 12.1 mmol) in anhydrous dichloromethane
(80 mL) was prepared and 1,8-bis(dimethylamino)naphthalene (7.78 g,
36.3 mmol) was added followed by trimethyloxonium tetrafluoroborate
(3.6 g, 24.2 mmol). The reaction mixture was stirred at RT for 3 h.
The reaction mixture was treated with methanol (3 mL) and stirred
for 5 min before adding hydrochloric acid (2 M, 200 mL) and ethyl
acetate (250 mL). The mixture was filtered to remove an insoluble
solid and the layers were separated. The aqueous layer was
extracted with ethyl acetate (2.times.100 mL). The organic layers
were combined and washed with brine, dried over sodium sulfate,
filtered and evaporated to give the title compound (4.80 g, 100%)
as a pale brown solid.
##STR00072##
A solution of lithium hydroxide in water (2 M, 50 mL, 100 mmol) was
added to a stirred solution of 5h (4.80 g, 12.1 mmol) in
tetrahydrofuran (130 mL). The reaction mixture was heated to
60.degree. C. for 15 h. The reaction mixture was cooled to RT,
before partially evaporating and adding hydrochloric acid (2 M, 150
mL). The mixture was extracted with ethyl acetate (3.times.50 mL).
The extract was dried over sodium sulfate, filtered and evaporated
to give a brown oil (3.5 g). The oil was purified by silica gel
chromatography using iso-hexanes/diethyl ether 1:1 to give the
title compound (1.132 g, 59%) as a colorless oil.
##STR00073##
A cooled (0.degree. C.) solution of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid 2,2,2-trichloro-ethyl
ester (2.174 g, 4.088 mmol) in anhydrous dichloromethane (50 mL)
was treated with trimethylsilyl trifluoromethanesulfonate (1.2 mL,
6.814 mmol). After 1 h at 0.degree. C., the reaction mixture was
treated with N,N-diisopropylethylamine (2.4 mL, 13.628 mmol) and
the volatiles were removed in vacuo to afford the corresponding
amine as a yellow foam. To this amine was added 5i (539.0 mg, 3.407
mmol), N,N-diisopropylethylamine (2.4 mL, 13.628 mmol) and
acetonitrile (50 mL). The reaction mixture was cooled to 0.degree.
C. and treated with
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (1.814 g, 4.770 mmol). After
overnight stirring at RT the reaction was quenched with 1 M
hydrochloric acid (100 mL). The aqueous layer was extracted with
ethyl acetate (2.times.). The organics were combined, dried over
sodium sulfate, filtered and the volatiles were removed in vacuo.
The residue was purified by silica gel chromatography using a 50 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 1:4 to afford the title compound
(2.193 g, 93%) as a light yellow solid.
##STR00074##
A solution of 5j (763.4 mg, 1.335 mmol) in tetrahydrofuran (25 mL)
was subsequently treated with zinc powder (1.920 g, 29.365 mmol)
and a solution of ammonium acetate (1.543 g, 20.025 mmol) in water
(5 mL). After overnight stirring the reaction was filtered through
Celite and quenched with 2 M hydrochloric acid. The aqueous layer
was extracted with ethyl acetate (2.times.). The organics were
combined, dried over sodium sulfate, filtered and the volatiles
were removed in vacuo. Residual acetic acid was azeotroped off with
toluene to provide the title compound (566.4 mg, 96%) as a light
orange solid.
##STR00075##
A solution of 5k (246 mg, 0.6 mmol),
(R)-1-(3-vinyl-isoquinolin-6-yl)-ethylamine dihydrochloride (162
mg, 0.6 mmol) and N,N-diisopropylethylamine (387 mg, 0.52 mL, 3
mmol) in acetonitrile (20 mL) was stirred at RT under nitrogen.
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (320 mg, 0.84 mmol) was added and
the reaction mixture was stirred at RT for 4 h. The solvent was
evaporated. The residue was diluted with ethyl acetate and
saturated sodium hydrogen carbonate solution. A small amount of
methanol (5 mL) was added to the suspension to give two clear
layers. The organic layer was separated, washed with water and
brine, dried over anhydrous sodium sulfate, filtered and
evaporated. The residue was purified by silica gel chromatography
using methanol/dichloromethane 1:20. The residue was triturated
with ether and the resulting solid was collected, washed with ether
and dried to afford the title compound (238 mg, 64%) as a pale
brown solid.
##STR00076##
A solution of 5l (91 mg, 0.15 mmol) in toluene (45 mL) was stirred
at RT under nitrogen. Hoveyda-Grubbs 2.sup.nd generation catalyst
(10 mg, 0.015 mmol) was added and the reaction mixture was heated
at reflux under nitrogen for 2 h. Additional Hoveyda-Grubbs
2.sup.nd generation catalyst (10 mg, 0.015 mmol) was added and the
reaction mixture was heated at reflux under nitrogen for 6 h. The
reaction mixture was cooled to RT, silica gel was added and the
reaction mixture was evaporated. The residue was purified by silica
gel chromatography using a gradient of ethyl acetate/acetone 20:1
to 5:2. The residue was triturated with ether and the resulting
solid was collected, washed with ether/iso-hexanes (1:1) and dried
to afford the title compound (23 mg, 26%) as a white solid. .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta.0.99 (d, J=6.7 Hz, 3H), 1.04 (d,
J=6.7 Hz, 3H), 1.44 (d, J=7.4 Hz, 3H), 1.53 (d, J=6.9 Hz, 3H), 1.61
(d, J=6.9 Hz, 3H), 1.68-2.20 (m, 5H), 2.62-2.74 (m, 2H), 3.35-3.43
(m, 1H), 3.44 (s, 3H), 3.72 (d, J=12.5 Hz, 1H), 3.88-3.92 (m, 2H),
4.23 (dd, J=8.9, 6.0 Hz, 1H), 4.55-4.60 (m, 1H), 5.30-5.37 (m, 1H),
5.80-5.90 (m, 1H), 6.38 (d, J=8.9 Hz, 1H), 6.43 (d, J=7.6 Hz, 1H),
6.93 (d, J=16.3 Hz, 1H), 7.24 (dd, J=16.3, 8.7 Hz, 1H), 7.45 (d,
J=8.7 Hz, 1H), 7.56 (d, J=8.5 Hz, 1H), 7.69 (s, 1H), 7.93 (d, J=8.5
Hz, 1H), 9.17 (s, 1H). LCMS (m/z) 579.3 [M+H], Tr=1.40 min.
Example 6
##STR00077##
A solution of Compound 5 (11 mg, 0.019 mmol) in ethyl acetate (10
mL) containing 10% palladium on carbon (10 mg) was hydrogenated at
RT and pressure for 3 h. The reaction mixture was filtered through
filter aid and the filter pad was washed with ethyl acetate. The
filtrate was evaporated and the residue was purified by silica gel
chromatography using a gradient of ethyl acetate/acetone 10:1 to
2:1. The residue was triturated with ethyl acetate/ether (1:5) to
afford the title compound (2.0 mg, 18%) as a white solid. .sup.1H
NMR (300 MHz, CDCl.sub.3) 0.93 (d, J=6.7 Hz, 3H), 0.98 (d, J=6.7
Hz, 3H), 1.39 (d, J=6.9 Hz, 3H), 1.55-2.70 (m, 8H), 1.95-2.50 (m,
5H), 2.64-2.73 (m, 2H), 2.95-3.08 (m, 2H), 3.35-3.42 (m, 2H), 3.53
(s, 3H), 3.61 (d, J=12.1 Hz, 1H), 4.06-4.11 (m, 1H), 4.50-4.57 (m,
1H), 5.25-5.32 (m, 1H), 5.64-5.70 (m, 1H), 6.39 (d, J=8.3 Hz, 1H),
6.47-6.51 (m, 1H), 7.21-7.28 (m, 1H), 7.41 (d, J=8.8 Hz, 1H), 7.62
(s, 1H), 7.69 (s, 1H), 7.92 (d, J=8.8 Hz, 1H), 9.15 (s, 1H). LCMS
(m/z) 581.3 [M+H], Tr=1.38 min.
Example 7
##STR00078##
A mixture of 7-bromo-2-chloro-quinoxaline (1.46 g, 6.00 mmol) and
tributyl(1-ethoxyvinyl)tin (2.71 g, 2.54 mL, 7.50 mmol) in toluene
(21 mL) was degassed for 20 min.
Bis(triphenylphosphine)palladium(II) dichloride (427 mg, 0.60 mmol)
was added and the reaction mixture stirred under nitrogen and
heated at 80.degree. C. for 19 h before allowing to cool. The
volatiles were evaporated and the residue suspended in 1,4-dioxane
(15 mL), 2 M aqueous hydrochloric acid (15 mL) was added and the
reaction mixture stirred for 45 min and then evaporated to remove
the volatiles. The residue was diluted with water and extracted
with ethyl acetate (2.times.) and the combined organic extracts
were washed with brine, dried over magnesium sulfate, filtered and
evaporated. The product was purified by chromatography using silica
gel doped with 10% w/w potassium carbonate eluting using a gradient
of iso-hexanes/ethyl acetate 1:0 to 9:1 to afford the title
compound (836 mg, 56%) as a yellow solid.
##STR00079##
Dichloro (p-cymene) ruthenium(II) dimer (12 mg, 0.019 mmol) and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (17 mg, 0.045
mmol) were suspended in degassed water (7.5 mL) and the mixture was
degassed with nitrogen for 15 min. The mixture was stirred at
70.degree. C. under nitrogen for 90 min. The resulting turbid
orange mixture was allowed to cool to RT. Solid 7a (948 mg, 3.78
mmol) followed by degassed tetrahydrofuran (7.5 mL) and sodium
formate (1.29 g, 18.9 mmol) were added and the reaction mixture was
degassed with nitrogen for 5 min. The reaction mixture was
vigorously stirred at 40.degree. C. for 3 h and allowed to cool. It
was then diluted with ethyl acetate and the mixture washed with
water (2.times.). The aqueous washes were back-extracted with ethyl
acetate and the combined organics washed with brine, dried over
magnesium sulfate, filtered and evaporated. The residue was
purified by silica gel chromatography eluting with
iso-hexanes/ethyl acetate 2:1 to afford the title compound (814 mg,
85%) as a purple solid.
##STR00080##
To a mixture of 7b (490 mg, 1.94 mmol), N,N-dicyclohexylmethylamine
(416 mg, 457 .mu.L, 2.13 mmol) and tert-butyl 3-butenoate (648 mg,
739 .mu.L, 4.56 mmol) in toluene (19 mL) was added
bis(tri-tert-butylphosphine)palladium(0) (41 mg, 0.080 mmol) under
nitrogen and the reaction mixture stirred and heated under reflux
for 5 h then allowed to cool. The mixture was evaporated and then
purified by silica gel chromatography using a gradient of
iso-hexanes/ethyl acetate 4:1 to 3:2 to afford the title compound
(367 mg, 60%) as a yellow oil.
##STR00081##
A solution of 1e (804 mg, 1.51 mmol) in tetrahydrofuran (37.7 mL)
was prepared and zinc powder (2.18 g, 33.3 mmol) was added followed
by a solution of ammonium acetate (1.75 g, 22.7 mmol) in water (9.4
mL). The reaction mixture was stirred at RT for 72 h. The reaction
was filtered through hyflo-supercel washing through with ethyl
acetate and saturated aqueous potassium hydrogen sulfate. The
mixture was treated with 1 M hydrochloric acid (3 mL) and the
layers were separated. The aqueous layer was extracted with ethyl
acetate (2.times.100 mL). The organic layers were combined, washed
with brine, filtered and evaporated to give a colorless gum. The
residue was azeotroped with toluene (3.times.200 mL) to give the
title compound (605 mg, quantitative yield) as a white solid.
##STR00082##
To a stirred solution of 7d (456 mg, 1.14 mmol) and
(E)-4-[3-((R)-1-hydroxy-ethyl)-quinoxalin-6-yl]-but-3-enoic acid
tert-butyl ester (358 mg, 1.14 mmol) in dichloromethane (22 mL) was
added 1-hydroxybenzotriazole containing approx. 20% water (270 mg,
1.60 mmol) followed by 4-dimethylaminopyridine (139 mg, 1.14 mmol)
and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(306 mg, 1.60 mmol). The reaction was stirred for 18 h and then
diluted with dichloromethane, washed with saturated ammonium
chloride solution (2.times.), dried over sodium sulfate, filtered
and evaporated. The residue was purified by silica gel
chromatography using a gradient of iso-hexanes/ethyl acetate 3:1 to
0:1 to afford the title compound (335 mg, 45%) as a white foam.
##STR00083##
To a stirred solution of 7e (309 mg, 0.444 mmol) in dichloromethane
(4.5 mL) at 0.degree. C. under nitrogen was added trimethylsilyl
trifluoromethanesulfonate (346 mg, 359 .mu.L, 1.56 mmol) and the
reaction mixture was allowed to warm to RT over 2.5 h.
N,N-diisopropylethylamine (164 mg, 221 .mu.L, 1.27 mmol) was added
and the reaction mixture stirred for a further 10 min, evaporated
and then suspended in acetonitrile (45 mL). The stirred mixture was
cooled to 0.degree. C. and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (236 mg, 0.622 mmol) and
N,N-diisopropylethylamine (229 mg, 309 .mu.L, 1.77 mmol) added.
After 90 min the reaction was quenched with a saturated ammonium
chloride solution and the mixture evaporated to remove organic
volatiles. The residue was diluted with dichloromethane and the
organic layer separated and washed with saturated sodium
bicarbonate (2.times.) and brine then dried over sodium sulfate,
filtered and evaporated. The residue was purified by silica gel
chromatography eluting with a gradient of ethyl acetate/acetone 1:0
to 9:1. The residue was further purified by reverse phase
preparative HPLC using acetonitrile/water 3:7 to afford the title
compound (7.6 mg, 3% over 2 steps) as a white solid. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta.1.00 (d, J=6.7 Hz, 3H), 1.01 (d, J=6.7
Hz, 3H), 1.62 (d, J=7.1 Hz, 3H), 1.66-1.75 (m, 2H), 1.77 (d, J=6.9
Hz, 3H), 1.91-2.05 (m, 3H), 2.72-2.82 (m, 1H), 2.98-3.08 (m, 1H),
3.38-3.41 (m, 1H), 3.78-3.84 (m, 1H), 4.25 (d, J=10.5 Hz, 1H), 4.41
(br d, J=11.3 Hz, 1H), 5.68 (q, J=7.1 Hz, 1H), 6.09 (q, J=6.9 Hz,
1H), 6.47 (d, J=16.3 Hz, 1H), 6.55-6.63 (m, 1H), 7.68 (s, 1H), 7.99
(s, 2H), 8.84 (s, 1H). LCMS (m/z) 523.2 [M+H], Tr=1.75 min.
Examples 8 and 9
##STR00084##
Dichloro (p-cymene) ruthenium(II) dimer (3 mg, 0.005 mmol) and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (4.4 mg, 0.012
mmol) were suspended in degassed water (2 mL) and the mixture was
degassed with nitrogen for 15 min. The mixture was stirred at
70.degree. C. under nitrogen for 90 min. The resulting yellow
solution was cooled to RT. 1-(3-Chloro-isoquinolin-6-yl)-ethanone
(206 mg, 1 mmol), sodium formate (340 mg, 5 mmol) and degassed
tetrahydrofuran (1 mL) were added and the reaction mixture was
degassed with nitrogen for 5 min. The reaction mixture was
vigorously stirred at 40.degree. C. for 2.5 h. The reaction mixture
was cooled to RT and was extracted with ethyl acetate. The organic
layer was separated, washed with water and brine, dried over sodium
sulfate, filtered and evaporated. The residue was purified by
silica gel chromatography using a gradient of iso-hexanes/ethyl
acetate 4:1 to 2:1 to afford the title compound (193 mg, 92%) as a
white solid.
##STR00085##
1,4-Dioxane (5 mL) was degassed with nitrogen, 8a (208 mg, 1 mmol),
tributyl(vinyl)tin (951 mg, 0.9 mL, 3 mmol) and
bis(triphenylphosphine)palladium(II) dichloride (70 mg, 0.1 mmol)
were added and the reaction mixture was heated at 150.degree. C. in
a microwave reactor for 1 h. Additional tributyl(vinyl)tin (0.3 mL,
1 mmol) and bis(triphenylphosphine)palladium(II) dichloride (70 mg,
0.1 mmol) were added and the reaction mixture was heated at
150.degree. C. in a microwave reactor for 1 h. The reaction mixture
was cooled to RT and the mixture was filtered through filter aid
and the filter pad was washed with ethyl acetate. The filtrate was
evaporated and the residue was purified by silica gel
chromatography using a gradient of iso-hexanes/ethyl acetate 4:1 to
2:1 followed by silica gel chromatography using iso-hexanes/ethyl
acetate 3:1 to afford the title compound (100 mg, 50%) as a white
solid.
##STR00086##
A solution of 1e (10.6 g, 20 mmol) in dichloromethane (300 mL) was
stirred at 0.degree. C. under nitrogen. Trimethylsilyl
trifluoromethanesulfonate (6.66 g, 5.4 mL, 30 mmol) was added
dropwise and the reaction mixture was stirred at 0.degree. C. for 1
h. Cold saturated sodium hydrogen carbonate solution (200 mL) was
added and the reaction mixture was stirred at 0.degree. C. for 15
min. The organic layer was separated, washed with saturated sodium
hydrogen carbonate solution and brine, dried over sodium sulfate,
filtered and evaporated to afford
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahy-
dro-pyridazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (20
mmol), which was used crude in the next step. A solution of
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (20 mmol) in
acetonitrile (240 mL) was stirred at 0.degree. C. under nitrogen.
(E)-Pent-3-enoic acid (2.20 g, 2.2 mL, 22 mmol) and
1-hydroxybenzotriazole hydrate (3.82 g, 20 mmol, wetted with not
less than 20 wt. % water) and
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (5.38
g, 28 mmol) was added and the reaction mixture was stirred at
0.degree. C. for 15 min and then at RT for 20 h. The solvent was
evaporated and the residue was partitioned between ethyl acetate
and water. The organic extracts were combined, washed with water
and brine, dried over anhydrous sodium sulfate, filtered and
evaporated. The residue was purified by silica gel chromatography
using iso-hexanes/ethyl acetate 3:1 to neat ethyl acetate to afford
the title compound (9.0 g, 88%) as a white solid.
##STR00087##
A solution of 8c (9.0 g, 17.5 mmol) in tetrahydrofuran (300 mL) was
stirred at RT under nitrogen. Zinc powder (25.0 g, 385 mmol) was
added followed by a solution of ammonium acetate (20.2 g, 263 mmol)
in water (200 mL). The reaction mixture was stirred at RT under
nitrogen for 18 h. The reaction mixture was filtered through Celite
and the filter pad was washed with water (100 mL) and ethyl acetate
(200 mL). The organic layer was separated and the solvent was
evaporated to ca. 100 mL and the solution was extracted with water
(100 mL). The aqueous layers were combined, saturated ammonium
chloride solution (150 mL) was added and the solution was acidified
to pH 1 with 2 M aqueous hydrochloric acid. The solution was
extracted with ethyl acetate and the organic extracts were
combined, washed with brine, dried over anhydrous magnesium
sulfate, filtered and evaporated. The residue was co-evaporated
with toluene (3.times.) to afford
(S)-1-{(S)-2-[(S)-3-methyl-2-((E)-pent-3-enoylamino)-butyrylamino]-propio-
nyl}-hexahydro-pyridazine-3-carboxylic acid (5.7 g, 85%) as a white
solid which was used in the next reaction. A solution of
(S)-1-{(S)-2-[(S)-3-methyl-2-((E)-pent-3-enoylamino)-butyrylamino]-propio-
nyl}-hexahydro-pyridazine-3-carboxylic acid (5.16 g, 13.5 mmol) in
dichloromethane (280 mL) and tetrahydrofuran (20 mL) was stirred at
RT under nitrogen. (R)-1-(3-Vinyl-isoquinolin-6-yl)-ethanol (2.69
g, 13.5 mmol) was added followed by
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (3.63
g, 18.9 mmol) and 4-(dimethylamino)pyridine (1.64 g, 13.5 mmol) and
the reaction mixture was stirred at RT for 6 h. Dichloromethane
(200 mL) was added and the solution was washed with aqueous citric
acid solution (pH 2-3). The organic layer was separated, washed
with water and brine, dried over anhydrous sodium sulfate, filtered
and evaporated. The residue was purified by silica gel
chromatography using a gradient of iso-hexanes/ethyl acetate 3:1 to
neat ethyl acetate followed by silica gel chromatography using
iso-hexanes/ethyl acetate 1:8 to afford the title compound (3.91 g,
51%) as a white solid.
##STR00088##
A solution of 8d (1.13 g, 2 mmol) in toluene (600 mL) was stirred
at RT under nitrogen for 15 min. Hoveyda-Grubbs 2.sup.nd generation
catalyst (125 mg, 0.2 mmol) was added and the reaction mixture was
heated at reflux under nitrogen for 30 min. Additional
Hoveyda-Grubbs 2.sup.nd generation catalyst (125 mg, 0.2 mmol) was
added and the reaction mixture was heated at reflux for 30 min. The
reaction mixture was cooled to RT. The majority of the solvent was
evaporated, silica gel was added and the solvent was evaporated.
The residue was purified by silica gel chromatography using a
gradient of ethyl acetate/acetone 9:1 to 3:2 followed by silica gel
chromatography using ethyl acetate/methanol 40:1. The residue was
triturated with ethyl acetate/ether (1:4) and the resulting solid
was collected, washed with ethyl acetate/ether (1:4) and dried to
afford a .about.10:1 mixture of Compound 9 and Compound 8, as a
pale brown solid (245 mg). A sample of the mixture was purified by
reverse phase preparative HPLC to afford Compound 8 (4 mg) as a
white solid. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.1.01 (d,
J=6.5 Hz, 3H), 1.02 (d, J=6.5 Hz, 3H), 1.37 (d, J=7.1 Hz, 3H), 1.65
(d, J=6.5 Hz, 3H), 1.70-2.15 (m, 5H), 2.95-3.05 (m, 1H), 3.70-3.76
(m, 1H), 4.03-4.10 (m, 2H), 4.12 (d, J=8.3 Hz, 1H), 5.64 (q, J=7.1
Hz, 1H), 6.10-6.20 (m, 2H), 6.98 (d, J=11.3 Hz, 1H), 7.65 (d, J=8.5
Hz, 1H), 7.93 (s, 1H), 8.05-8.10 (m, 2H), 9.21 (s, 1H). LCMS (m/z)
522.3 [M+H], Tr=1.25 min.
Compound 9, .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.0.97 (d, J=6.9
Hz, 3H), 0.99 (d, J=6.7 Hz, 3H), 1.53 (d, J=6.9 Hz, 3H), 1.71 (d,
J=6.9 Hz, 3H), 1.75-2.70 (m, 6H), 3.20-3.40 (m, 2H), 3.63-3.77 (m,
2H), 4.23-4.29 (m, 1H), 4.53-5.57 (m, 1H), 5.65-5.76 (m, 1H), 6.04
(q, J=6.7 Hz, 1H), 6.38-6.53 (m, 3H), 6.72 (d, J=16.3 Hz, 1H), 7.39
(d, J=8.3 Hz, 1H), 7.55 (s, 1H), 7.72 (s, 1H), 7.92 (d, J=8.3 Hz,
1H), 9.13 (s, 1H). LCMS (m/z) 522.0 [M+H], Tr=1.40 min.
Example 10
##STR00089##
10a was prepared in the same manner as 4d using 3-butenoic acid
instead of 4,4-dimethyl-hept-6-enoic acid in 66% yield.
##STR00090##
10b was prepared in the same manner as 4e using
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester
instead of
(S)-1-{(S)-2-[(S)-2-(4,4-dimethyl-hept-6-enoyloxy)-3-methyl-butyrylami-
no]-propionyl}-hexahydro-pyridazine-3-carboxylic acid
2,2,2-trichloro-ethyl ester in 84% yield.
##STR00091##
10c was prepared in the same manner as 8d using
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid instead of
(S)-1-{(S)-2-[(S)-3-methyl-2-((E)-pent-3-enoylamino)butyrylamino]-propion-
yl}-hexahydro-pyridazine-3-carboxylic acid in 84% yield.
##STR00092##
Compound 9 was prepared in the same manner as Compound 5 using
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid
(R)-1-(3-vinyl-isoquinolin-6-yl)-ethyl ester instead of
(S)-1-{(S)-2-[(S)-2-((E)-(2R,3R)-3-methoxy-2-methyl-hex-4-enoylamino)-3-m-
ethyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylic
acid [(R)-1-(3-vinyl-isoquinolin-6-yl)-ethyl]-amide in 9%
yield.
##STR00093##
A solution of Compound 9 (16 mg, 0.03 mmol) in ethyl acetate (5 mL)
and tetrahydrofuran (2 mL) containing 10% palladium on carbon (15
mg) was hydrogenated at RT and pressure for 3 h. The reaction
mixture was filtered through filter aid and the filter pad was
washed with ethyl acetate. The filtrate was evaporated and the
residue was triturated with ether (2.times.2 mL) and the resulting
solid was dried to afford the title compound (7.6 mg, 48%) as an
off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.0.90 (d,
J=6.7 Hz, 3H), 0.95 (d, J=6.5 Hz, 3H), 1.50 (d, J=7.1 Hz, 3H), 1.68
(d, J=6.5 Hz, 3H), 1.60-2.40 (m, 9H), 2.70-3.10 (m, 3H), 3.68-3.75
(m, 1H), 3.90 (d, J=11.8 Hz, 1H), 4.32-4.37 (m, 1H), 4.44-4.49 (m,
1H), 5.83-5.92 (m, 1H), 6.05-6.11 (m, 2H), 6.33 (d, J=7.8 Hz, 1H),
7.34 (s, 1H), 7.36 (d, J=8.3 Hz, 1H), 7.80 (s, 1H), 7.93 (d, J=8.3
Hz, 1H), 9.17 (s, 1H). LCMS (m/z) 524.3 [M+H], Tr=0.59 min.
Examples 11 and 12
##STR00094##
To a stirred solution of 1e (10.6 g, 20.0 mmol) in dichloromethane
(400 mL), at 0.degree. C. under nitrogen, was added trimethylsilyl
trifluoromethanesulfonate (6.67 g, 5.43 mL, 30.0 mmol) and the
reaction mixture stirred at 0.degree. C. for 2 h.
N,N-Diisopropylethylamine (10.3 g, 13.9 mL, 80.0 mmol) was added
and the mixture allowed to warm to ambient temperature. The
volatiles were evaporated and the residue suspended in acetonitrile
(250 mL). The stirred mixture was cooled to 0.degree. C. under a
blanket of nitrogen and then N,N-diisopropylethylamine (10.3 g,
13.9 mL, 80 mmol) and 3-butenoic acid (1.89 g, 1.86 mL, 4.40 mmol)
added, followed by
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (10.6 g, 28.0 mmol), portionwise.
The reaction mixture was allowed to warm to ambient temperature and
stirred for 18 h before evaporating. The residue was diluted with
ethyl acetate and then washed successively with saturated sodium
bicarbonate solution, water, 2 M hydrochloric acid, water then
brine, dried over magnesium sulfate, filtered and evaporated. The
residue was purified by silica gel chromatography using a gradient
of iso-hexanes/ethyl acetate 1:1 to 0:1 to afford a .about.1:1
mixture of
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester
and
(S)-1-{(S)-2-[(S)-2-((E)-but-2-enoylamino)-3-methyl-butyrylamino]-propion-
yl}-hexahydro-pyridazine-3-carboxylic acid 2,2,2-trichloro-ethyl
ester as an orange foam (7.23 g, 72%).
To a stirred solution of a mixture of
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester
and
(S)-1-{(S)-2-[(S)-2-((E)-but-2-enoylamino)-3-methyl-butyrylamino]-propion-
yl}-hexahydro-pyridazine-3-carboxylic acid 2,2,2-trichloro-ethyl
ester (.about.1:1, 0.99 g, 2.00 mmol) in tetrahydrofuran (40 mL)
was added zinc powder (2.86 g, 44.0 mmol) and a solution of
ammonium acetate (2.31 g, 30.0 mmol) in water (25 mL). The reaction
mixture was allowed to warm to ambient temperature and stirred for
18 h, then diluted with ethyl acetate and the mixture filtered.
From the filtrate the aqueous layer was separated and diluted with
an equal amount of saturated ammonium chloride solution and then
acidified to pH 1 with 2 M hydrochloric acid. The aqueous layer was
extracted with ethyl acetate (2.times.) and the combined organic
layers washed with brine, dried over magnesium sulfate, filtered
and evaporated. The residue was azeotroped with toluene (3.times.)
to give a mixture of the title compounds (.about.1:1, 466 mg, 63%)
as a yellow foam.
##STR00095##
A mixture of 11a and 12b (.about.1:1, 250 mg, 0.68 mmol) in
acetonitrile (20 mL) was stirred at RT under nitrogen.
(R)-1-(3-vinyl-isoquinolin-6-yl)-ethylamine dihydrochloride (136
mg, 0.5 mmol) and N,N-diisopropylethylamine (323 mg, 0.44 mL, 2.5
mmol) was added followed by
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (266 mg, 0.7 mmol) and the
reaction mixture was stirred at RT for 18 h. The solvent was
evaporated. The residue was diluted with ethyl acetate and the
solution was washed with saturated sodium hydrogen carbonate
solution, water and brine. The organic extract was dried over
anhydrous magnesium sulfate, filtered and evaporated. The residue
was purified by silica gel chromatography using a gradient of ethyl
acetate to methanol/ethyl acetate 1:5 to afford a .about.1:1
mixture of
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid
[(R)-1-(3-vinyl-isoquinolin-6-yl)-ethyl]-amide and
(S)-1-{(S)-2-[(S)-2-((E)-but-2-enoylamino)-3-methyl-butyrylamino]-propion-
yl}-hexahydro-pyridazine-3-carboxylic acid
[(R)-1-(3-vinyl-isoquinolin-6-yl)-ethyl]-amide as a white solid
(153 mg, 56%) which was used in the next step without further
purification.
A solution of a mixture of
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid
[(R)-1-(3-vinyl-isoquinolin-6-yl)-ethyl]-amide and
(S)-1-{(S)-2-[(S)-2-((E)-but-2-enoylamino)-3-methyl-butyrylamino]-propion-
yl}-hexahydro-pyridazine-3-carboxylic acid
[(R)-1-(3-vinyl-isoquinolin-6-yl)-ethyl]-amide (.about.1:1, 150 mg,
0.27 mmol) in toluene (70 mL) was stirred at RT under nitrogen.
Hoveyda-Grubbs 2.sup.nd generation catalyst (17 mg, 0.027 mmol) was
added and the reaction mixture was heated at reflux under nitrogen
for 90 min. Additional Hoveyda-Grubbs 2.sup.nd generation catalyst
(17 mg, 0.027 mmol) was added and the reaction mixture was heated
at reflux under nitrogen for 90 min. The reaction mixture was
cooled to RT, silica gel was added and the solvent was evaporated.
The residue was purified by silica gel chromatography using a
gradient of ethyl acetate to ethyl acetate/acetone 1:1. The residue
was further purified by preparative thin layer chromatography using
methanol/ethyl acetate 1:4 followed by silica gel chromatography
using dichloromethane/methanol 20:1 to give the title compounds.
Compound 11 (1.3 mg, 1%) as a white solid: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta.1.01 (d, J=6.7 Hz, 3H), 1.02 (d, J=6.7 Hz, 3H),
1.61 (d, J=7.1 Hz, 3H), 1.66 (d, J=7.1 Hz, 3H), 1.80-2.05 (m, 5H),
2.71-2.82 (m, 1H), 2.99-3.05 (m, 1H), 3.32-3.43 (m, 1H), 3.64-3.75
(m, 1H), 4.27 (d, J=10.1 Hz, 1H), 4.40-4.53 (m, 2H), 5.12 (q, J=7.1
Hz, 1H), 5.62 (q, J=7.1 Hz, 1H), 6.47-6.56 (m, 1H), 6.62 (d, J=15.8
Hz, 1H), 7.43 (s, 1H), 7.61 (dd, J=8.5, 1.6 Hz 1H), 7.74 (s, 1H),
8.05 (d, J=8.5 Hz, 1H), 9.13 (s, 1H). LCMS (m/z) 521.3 [M+H],
Tr=0.97 min.
Compound 12 (1.7 mg, 1.2%) as a white solid: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta.0.98 (d, J=6.7 Hz, 3H), 1.03 (d, J=6.7 Hz, 3H),
1.47 (d, J=7.1 Hz, 3H), 1.60 (d, J=6.9 Hz, 3H), 1.65-2.16 (m, 5H),
2.81-2.89 (m, 1H), 3.61-3.69 (m, 1H), 4.04 (d, J=6.9 Hz, 1H),
4.30-4.36 (m, 1H), 4.75 (d, J=11.6 Hz, 1H), 5.23 (q, J=7.1 Hz, 1H),
5.90 (q, J=6.9 Hz, 1H), 6.36 (d, J=13.1 Hz, 1H), 6.92 (d, J=13.1
Hz, 1H), 7.65 (d, J=8.5 Hz 1H), 7.88 (s, 1H), 8.05 (d, J=8.5 Hz
1H), 8.46 (s, 1H), 9.16 (s, 1H). LCMS (m/z) 507.2 [M+H], Tr=0.95
min.
Example 13
##STR00096##
A suspension of
(S)-1-(7-bromo-quinolin-2-yl)-2,2,2-trifluoro-ethylamine
hydrochloride (Asiba Pharmatech, Edison, N.J., USA, 397 mg, 1.16
mmol) in dichloromethane (10 mL) was cooled using an ice bath.
Triethylamine (985 .mu.L, 3.48 mmol) was added and the reaction was
stirred until a homogenous solution was observed. Di-tert-butyl
dicarbonate (380 mg, 1.74 mmol) in dichloromethane (5 mL) was then
added and the reaction was left to stir overnight. Di-tert-butyl
dicarbonate (127 mg, 0.58 mmol) was added and the reaction was
stirred for a further 6 h. Di-tert-butyl dicarbonate (253 mg, 1.16
mmol) was added along with 4 .ANG. molecular sieves and the
reaction was left to stir overnight. The reaction was washed with
water and brine and the organics were dried over anhydrous sodium
sulfate, filtered and concentrated. The residue was purified by
silica gel chromatography using a stepwise gradient from 1:0 to 7:3
iso-hexanes/ethyl acetate to afford the title compound (343 mg,
73%) as an orange solid.
##STR00097##
To a 3-necked flask, fitted with stirrer bar and thermometer, under
an atmosphere of nitrogen and containing magnesium turnings (3.02
g, 0.124 mol) was added anhydrous tetrahydrofuran (105 mL). A
single crystal of iodine was added to the reaction followed by
1-chloro-3-methyl-but-2-ene (2 mL, 17.75 mmol). The reaction was
stirred for 20 min until all color had left the solution.
1-chloro-3-methyl-but-2-ene (5.57 mL, 49.45 mmol) was added. A
gradual temperature increase was observed until the reaction had
reached reflux. The reaction was left to stir for 1 hour, allowing
the reaction to return to RT. The reaction was then transferred via
cannula to a flask containing solid carbon dioxide (50 g). The
reaction was then stirred for 2 h. A cooling ice bath was added and
the reaction was quenched with 2 M hydrochloric acid. The reaction
mixture was then extracted with diethyl ether. The organics were
then dried over anhydrous sodium sulfate, filtered and
concentrated. The residue was purified by silica gel chromatography
using a stepwise gradient from 1:0 to 9:1 iso-hexanes/ethyl acetate
to afford the title compound (2.69 g, 35%) as a pale yellow oil.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta.1.35 (s, 6H), 5.10-5.21
(m, 2H), 6.07 (dd, J=17.6, 10.5 Hz, 1H).
##STR00098##
13a (343 mg, 0.846 mmol) and 13b (106 mg, 0.931 mmol) were placed
in a microwave vessel and dissolved in acetonitrile (3 mL).
Palladium(II) acetate (19 mg, 0.0846 mmol), tri(o-tolyl)phosphine
(51 mg, 0.169 mmol) and triethylamine (236 .mu.L, 1.69 mmol) were
added and the vessel was sealed before being irradiated in the
microwave for 30 min, using fixed hold time, on high absorption at
100.degree. C. The solvent was removed and the residue was taken up
in a mixture of water and ethyl acetate. The phases were separated
and the aqueous was extracted with ethyl acetate. The combined
organics were then dried over anhydrous sodium sulfate, filtered
and concentrated. The residue was purified by silica gel
chromatography using a stepwise gradient of iso-hexanes/ethyl
acetate from 1:0 to 6:4. The isolated material was subjected to a
second purification using the same conditions to afford the title
compound (142 mg, 38%) as a pale yellow oil.
##STR00099##
(S)-1-[(S)-2-((S)-2-Amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid ethyl ester (210 mg, 0.486 mmol) and 13c
(142 mg, 0.324 mmol) were dissolved in N,N-dimethylformamide (5
mL), under an atmosphere of nitrogen, and cooled using an ice bath.
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (203 mg, 0.535 mmol) and
N,N-diisopropylethylamine (423 .mu.L, 2.43 mmol) were then added
and the reaction was allowed to slowly warm to RT and left to stir
overnight. The reaction was diluted with water and extracted with
ethyl acetate. The combined organics were dried over anhydrous
sodium sulfate, filtered and concentrated. The residue was purified
by silica gel chromatography using a stepwise gradient of
iso-hexanes/ethyl acetate from 1:0 to 0:1 to afford the product
contaminated with residual solvent. Toluene was added and the
solution was concentrated to afford the title compound (79 mg, 29%)
as an orange solid.
##STR00100##
13d (79 mg, 0.0925 mmol) was dissolved in tetrahydrofuran (2 mL)
and cooled using an ice bath. Methanol (1 mL) and water (1 mL) were
then added followed by lithium hydroxide monohydrate (15 mg, 0.37
mmol). The reaction was then left to stir for 1 hour. 1 M
hydrochloric acid was added until the solution was pH 2. The
solvent was removed and the resultant solid was sequentially
azeotroped with methanol, then acetonitrile and finally toluene to
afford the title compound (67 mg, 100%) as a yellow solid.
##STR00101##
13e (66 mg, 0.0916 mmol) was dissolved in 4 M hydrochloric acid in
1,4-dioxane (2 mL) and left to stir for 30 min. The solvent was
removed and the resultant solid was triturated with diethyl ether
and dried to afford the title compound (50 mg, 83%) as a pale
yellow solid.
##STR00102##
13f (50 mg, 0.0761 mmol) was dissolved in dichloromethane (76 mL),
under an atmosphere of nitrogen and cooled using an ice bath.
2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (58 mg, 0.152 mmol) and
N,N-diisopropylethylamine (53 .mu.L, 0.304 mmol) were then added
and the reaction was allowed to slowly warm to RT and left to stir
overnight. The solvent was removed and the residue was purified by
silica gel chromatography using 7:3 iso-hexanes/acetone. The
residue was re-purified by silica gel chromatography using a
stepwise gradient of iso-hexanes/acetone from 1:0 to 1:1. The
residue was then eluted through an HPLC system fitted with a
Phenomenex Gemini 10.mu. 110 A, 250.times.21.2 mm column using a
continuous gradient of acetonitrile/water from 1:4 to 1:0 flow at
20 mL/min to afford the title compound (7 mg, 15%) as a solid.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta.0.86-1.08 (m, 6H),
1.25-1.54 (m, 8H), 1.57-1.81 (m, 5H), 1.88-2.00 (m, 1H), 2.37-2.80
(m, 1H), 3.32-3.65 (m, 1H), 3.69-3.97 (m, 1H), 4.22-4.39 (m, 1H),
4.50-4.68 (m, 1H), 5.74-6.05 (m, 2H), 6.22-6.39 (m, 1H), 6.44-6.54
(m, 1H), 6.63-6.76 (m, 1H), 7.45 (app t, J=8.5 Hz, 1H), 7.56 (app
t, J=7.6 Hz, 1H), 7.81 (d, J=8.3 Hz, 1H), 8.19 (s, 1H), 8.76-9.09
(m, 1H). LCMS (m/z) 603.1 [M+H], Tr=2.59 min.
Example 14
##STR00103##
Dichloro (p-cymene) ruthenium(II) dimer (24 mg, 0.040 mmol) and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (35 mg, 0.096
mmol) were suspended in degassed water (16 mL) and the mixture was
degassed with nitrogen for 15 min. The mixture was stirred at
70.degree. C. under nitrogen for 90 min. The resulting turbid
orange mixture was allowed to cool to RT. Solid
1-(6-bromo-quinolin-3-yl)-ethanone (prepared as in WO2011/063233,
1.92 g, 7.68 mmol) followed by degassed tetrahydrofuran (16 mL) and
sodium formate (2.72 g, 40 mmol) were added and the reaction
mixture was degassed with nitrogen for 5 min, further degassed
tetrahydrofuran (5 mL) was added and the mixture degassed for
another minute. The reaction mixture was vigorously stirred at
40.degree. C. for 21 h and allowed to cool. It was then diluted
with ethyl acetate and the mixture washed with water then brine,
dried over magnesium sulfate, filtered and evaporated. The residue
was purified by silica gel chromatography using a gradient of
iso-hexanes/ethyl acetate 7:3 to 0:1 to afford the title compound
(1.65 g, 85%) as a brown solid.
##STR00104##
To a mixture of 14a (356 mg, 1.41 mmol),
N,N-dicyclohexylmethylamine (275 mg, 301 .mu.L, 1.41 mmol) and
tert-butyl 3-butenoate (470 mg, 537 .mu.L, 3.31 mmol) in toluene
(14 mL) was added bis(tri-tert-butylphosphine)palladium(0) (30 mg,
0.058 mmol) under nitrogen and the reaction mixture stirred and
heated under reflux for 90 min then allowed to cool. The mixture
was evaporated and then purified by silica gel chromatography using
a gradient of iso-hexanes/ethyl acetate 4:1 to 2:3 to afford the
title compound as a yellow oil (144 mg, 33%).
##STR00105##
To a stirred solution of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid (189 mg, 0.473 mmol)
and 14b (148 mg, 0.473 mmol) in dichloromethane (10 mL) was added
1-hydroxybenzotriazole containing approx. 20% water (89 mg, 0.662
mmol) followed by 4-dimethylaminopyridine (58 mg, 0.473 mmol) then
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (127
mg, 0.662 mmol). The reaction was stirred for 62 h and then diluted
with dichloromethane, washed with saturated ammonium chloride
solution (2.times.) and brine, dried over magnesium sulfate,
filtered and evaporated. Purified by silica gel chromatography
using a gradient of iso-hexanes/ethyl acetate 3:1 to 2:3 afforded
the title compound (126 mg, 38%) as a white foam.
##STR00106##
To a stirred solution of 14c (126 mg, 0.181 mmol) in
dichloromethane (1.8 mL) at 0.degree. C. under nitrogen was added
trimethylsilyl trifluoromethanesulfonate (141 mg, 147 .mu.L, 0.635
mmol) and the reaction mixture was allowed to warm to ambient
temperature over 1.75 h. Further trimethylsilyl
trifluoromethanesulfonate (20 mg, 21 .mu.L, 0.091 mmol) was added
and the reaction mixture was stirred for a further 15 min before
the addition of N,N-diisopropylethylamine (164 mg, 221 .mu.L, 1.27
mmol). After a further 10 min of stirring the reaction mixture was
evaporated and then suspended in acetonitrile (18.1 mL). The
stirred mixture was cooled to 0.degree. C. and then
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (96 mg, 0.253 mmol) and
N,N-diisopropylethylamine (94 mg, 126 .mu.L, 0.724 mmol) added.
After 45 min the reaction was quenched with saturated ammonium
chloride solution (10 mL) and the mixture evaporated to remove
organic volatiles. The residue was diluted with dichloromethane and
the aqueous layer separated and extracted with dichloromethane. The
organic extracts were washed with saturated sodium bicarbonate,
water and then brine, dried over sodium sulfate, filtered and
evaporated. The residue was purified by silica gel chromatography
eluting with ethyl acetate/acetone 9:1 then by preparative reverse
phase HPLC using a gradient of acetonitrile/water 3:7 to 1:1 to
afford the title compound (6.5 mg, 7% over 2 steps) as a white
solid. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.1.00 (d, J=6.7 Hz,
3H), 1.03 (d, J=6.7 Hz, 3H), 1.61 (d, J=7.1 Hz, 3H), 1.75 (d, J=6.7
Hz, 3H), 1.91-2.01 (m, 3H), 2.68-2.79 (m, 1H), 3.00 (dd, J=14.5,
3.6 Hz, 1H), 3.35-3.42 (m, 1H), 3.73-3.82 (m, 1H), 4.29 (d, J=9.6
Hz, 1H), 4.41 (d, J=12.9 Hz, 1H), 4.70 (d, J=12.3 Hz, 1H), 5.47 (q,
J=7.3 Hz, 1H), 6.15 (q, J=6.6 Hz, 1H), 6.30-6.40 (m, 1H), 6.63 (d,
J=16.1 Hz, 1H), 7.55 (s, 1H), 7.82 (dd, J=8.7, 1.5 Hz, 1H), 7.95
(d, J=8.7 Hz, 1H), 8.21 (s, 1H), 8.79 (d, J=1.8 Hz, 1H). LCMS (m/z)
522.2 [M+H], Tr=1.60 min.
Example 15
##STR00107##
To a stirred mixture of 3-bromo-quinoline-6-carboxylic acid
(prepared as in WO2011/090935, 1.94 g, 7.70 mmol) in
tetrahydrofuran (77 mL) was added
2-chloro-4,6-dimethoxy-1,3,5-triazine (2.03 g, 11.5 mmol) and
N-methylmorpholine (2.34 g, 2.54 mL, 23.1 mmol). The reaction
mixture was stirred for 90 min and then N,O-dimethylhydroxylamine
hydrochloride (751 mg, 7.70 mmol) added in one portion. The
reaction mixture was stirred for a further 17 h and further
N,O-dimethylhydroxylamine hydrochloride (375 mg, 3.85 mmol) was
added and then after 5 h a further portion (175 mg, 1.80 mmol). The
reaction mixture was stirred for a further hour and then diluted
with dichloromethane, washed with water and then saturated ammonium
chloride solution. The organic layer was dried over sodium sulfate,
filtered and evaporated. The residue was purified by silica gel
chromatography eluting with a gradient of iso-hexanes/ethyl acetate
4:1 to 7:3 to afford the title compound (1.43 g, 66%) as an
off-white solid.
##STR00108##
To a stirred solution of methyl magnesium chloride (3 M in
tetrahydrofuran, 4.85 mL, 14.5 mmol) in tetrahydrofuran (10 mL)
under nitrogen was added a solution of 15a (1.43 g, 4.85 mmol) in
tetrahydrofuran (20 mL). The reaction mixture was stirred for 1
hour and then quenched with saturated ammonium chloride solution.
The mixture was diluted with diethyl ether and water, the organic
layer separated and washed with further water and the combined
aqueous washes back-extracted with diethyl ether. The combined
organic extracts were dried over magnesium sulfate, filtered and
evaporated. The residue was purified by silica gel chromatography
eluting with iso-hexanes/ethyl acetate 4:1 to afford the title
compound as a white solid which was taken directly into the next
step.
##STR00109##
Dichloro (p-cymene) ruthenium(II) dimer (9 mg, 0.015 mmol) and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (14 mg, 0.037
mmol) were suspended in degassed water (9 mL) and the mixture was
degassed with nitrogen for 15 min. The mixture was stirred at
70.degree. C. under nitrogen for 90 min. The resulting turbid
orange mixture was allowed to cool to RT. Solid
1-(3-bromo-quinolin-6-yl)-ethanone (771 mg, 3.08 mmol) and sodium
formate (2.72 g, 40 mmol) followed by degassed tetrahydrofuran (4.5
mL) were added and the reaction mixture was degassed with nitrogen
for 5 min. The reaction mixture was vigorously stirred at
40.degree. C. for 3 h and allowed to cool. It was then diluted with
ethyl acetate and the mixture washed with water then brine, dried
over sodium sulfate, filtered and evaporated. The residue was
purified by silica gel chromatography using a gradient of
iso-hexanes/ethyl acetate 3:2 to 1:1 to afford the title compound
(616 mg, 79%) as a grey solid.
##STR00110##
To a solution of 15c (560 mg, 2.22 mmol) in toluene (20 mL) and
1,4-dioxane (5 mL) was added N,N-dicyclohexylmethylamine (740
.mu.L, 3.46 mmol) and tert-butyl 3-butenoate (741 mg, 844 .mu.L,
5.22 mmol) was added bis(tri-tert-butylphosphine)palladium(0) (68
mg, 0.13 mmol) under nitrogen and the reaction mixture stirred and
heated under reflux for 6 h then allowed to cool. The mixture was
evaporated and then purified by silica gel chromatography using a
gradient of iso-hexanes/ethyl acetate 4:1 to 0:1 to give a yellow
gum. The gum was suspended in ethyl acetate and washed with
saturated ammonium chloride solution (2.times.) followed by water
then brine, dried over sodium sulfate, filtered and evaporated to
afford the title compound (300 mg, 43%) as a yellow oil.
##STR00111##
To a stirred slurry of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid (325 mg, 0.812 mmol),
15d (231 mg, 0.738 mmol) and powdered 4 .ANG. molecular sieves in
dichloromethane (16 mL) was added
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (198
mg, 1.03 mmol), 1-hydroxybenzotriazole hydrate containing approx.
20% water (174 mg, 1.03 mmol) followed by 4-dimethylaminopyridine
(90 mg, 0.738 mmol). The reaction was stirred under nitrogen for 16
h and then filtered and the solid washed with dichloromethane. The
filtrate was washed with saturated ammonium chloride solution
(2.times.) and water, dried over sodium sulfate, filtered and
evaporated. The residue was purified by silica gel chromatography
using a gradient of iso-hexanes/ethyl acetate 1:1 to 0:1 to afford
the title compound (307 mg, 60%) as a yellow foam.
##STR00112##
To a stirred solution of 15e (150 mg, 0.216 mmol) in
dichloromethane (2 mL) at 0.degree. C. under nitrogen was added
trimethylsilyl trifluoromethanesulfonate (168 mg, 174 .mu.L, 0.755
mmol) dropwise and the reaction mixture was allowed to warm to
ambient temperature over 2.25 h. N,N-Diisopropylethylamine (195 mg,
263 .mu.L, 1.51 mmol) was added and the reaction mixture stirred
for a further 10 min, evaporated and then suspended in acetonitrile
(21.6 mL). The stirred mixture was cooled to 0.degree. C. and then
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (115 mg, 0.320 mmol) and
N,N-diisopropylethylamine (111 mg, 150 .mu.L, 0.864 mmol) added.
After 1 h the reaction was quenched with saturated ammonium
chloride solution and the mixture evaporated to remove organic
volatiles. The residue was diluted with dichloromethane and the
organic layer separated and washed with saturated ammonium chloride
(2.times.) and brine then dried over sodium sulfate, filtered and
evaporated. The residue was purified by preparative reverse phase
HPLC using a gradient of acetonitrile/water 3:7 to 11:9 to afford
the title compound (4.6 mg, 4% over 2 steps) as an off-white solid.
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta.1.01 (app t, J=6.7 Hz,
6H), 1.62 (d, J=7.1 Hz, 3H), 1.70 (d, J=6.7 Hz, 3H), 1.75-1.85 (m,
1H), 1.91-2.05 (m, 3H), 2.70-2.80 (m, 1H), 2.98-3.06 (m, 1H),
3.36-3.45 (m, 1H), 3.74-3.84 (m, 1H), 4.26 (d, J=10.5 Hz, 1H), 4.42
(br d, J=12.3 Hz, 1H), 4.72-4.78 (m, 1H), 5.55 (q, J=7.1 Hz, 1H),
6.09 (q, J=6.5 Hz, 1H), 6.38-6.41 (m, 1H), 6.71 (d, J=16.1 Hz, 1H),
7.68 (dd, J=8.9, 1.9 Hz, 1H), 7.88-8.01 (m, 3H), 8.86 (d, J=2.0 Hz,
1H). LCMS (m/z) 522.3 [M+H], Tr=1.64 min.
Example 16
##STR00113##
A mixture of 7-bromo-2-chloro-quinoline (1.05 g, 4.32 mmol) and
tributyl(1-ethoxyvinyl)tin (1.95 g, 1.83 mL, 5.40 mmol) in toluene
(21 mL) was degassed for 20 min.
Bis(triphenylphosphine)palladium(II) dichloride (302 mg, 0.432
mmol) was added and the reaction mixture stirred under nitrogen and
heated at 80.degree. C. for 24 h before allowing to cool. The
volatiles were evaporated and the residue suspended in 1,4-dioxane
(10 mL) and 2 M aqueous hydrochloric acid (5 mL) was added and the
reaction mixture stirred for 30 min and then evaporated to remove
organics. The residue was diluted with ethyl acetate and water and
the organic layer washed with brine, dried over sodium sulfate,
filtered and evaporated. The product was purified on silica gel
doped with 10% w/w potassium carbonate eluting using a gradient of
iso-hexanes/ethyl acetate 9:1 to 4:1 to afford the title compound
(422 mg, 48%) as a yellow solid.
##STR00114##
Dichloro(p-cymene)ruthenium(II) dimer (8.5 mg, 0.014 mmol) and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (12.1 mg, 0.033
mmol) were suspended in degassed water (5.5 mL) and the mixture was
degassed with nitrogen for 20 min. The mixture was stirred at
70.degree. C. under nitrogen for 90 min. The resulting turbid
orange mixture was allowed to cool to RT. Solid 16a (571 mg, 2.78
mmol) and sodium formate (945 mg, 13.9 mmol) followed by degassed
tetrahydrofuran (5.5 mL) were added and the reaction mixture was
degassed with nitrogen for 5 min. The reaction mixture was
vigorously stirred at 40.degree. C. for 4 h and allowed to cool. It
was then diluted with ethyl acetate and the mixture washed with
water and the aqueous layer back-extracted with ethyl acetate. The
combined organic extracts were washed with brine, dried over sodium
sulfate, filtered and evaporated. The residue was purified by
silica gel chromatography using a gradient of iso-hexanes/ethyl
acetate 4:1 to 7:3 to afford the title compound (413 mg, 72%) as a
beige solid.
##STR00115##
To a mixture of 16b (360 mg, 1.73 mmol), potassium
vinyltrifluoroborate (279 mg, 2.08 mmol) and
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II),
complex with dichloromethane (27 mg, 0.033 mmol) in dry n-propanol
(27 mL) was added triethylamine (175 mg, 241 .mu.L, 1.73 mmol) and
the system evacuated and purged with nitrogen (3.times.). The
reaction mixture was stirred and heated at reflux for 3 h before
being allowed to cool. The mixture was poured into water and
extracted with diethyl ether (2.times.) and the combined organic
extracts dried over magnesium sulfate, filtered and evaporated. The
residue was purified by silica gel chromatography using a gradient
of iso-hexanes/ethyl acetate 7:3 to 3:2 to afford the title
compound (270 mg, 78%) as a white solid.
##STR00116##
To a stirred mixture of 16c (100 mg, 0.500 mmol) and
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid (203 mg, 0.550 mmol) in
dichloromethane (10 mL) under nitrogen was added
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (134
mg, 0.700 mmol) and 4-dimethylaminopyridine (61 mg, 0.500 mmol).
The reaction mixture was stirred for 16 h and then diluted with
dichloromethane, washed successively with citric acid solution,
water then brine, dried over sodium sulfate, filtered and
evaporated. The residue was purified by silica gel chromatography
using a gradient of iso-hexanes/ethyl acetate 1:1 to 0:1 to afford
the title compound (103 mg, 38%) as a brown solid.
##STR00117##
16d (60 mg, 0.109 mmol) and Hoveyda-Grubbs 2.sup.nd generation
catalyst (20.5 mg, 0.0328 mmol) was suspended in 1,2-dichloroethane
(6 mL) and the mixture heated in a microwave reactor at 100.degree.
C. for 1 h. The resulting mixture was combined with an identical
reaction carried out using (10 mg, 0.0182) of the
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid
(R)-1-(2-vinyl-quinolin-7-yl)-ethyl ester. The volatiles were
evaporated and the residue purified by silica gel chromatography
using a gradient of ethyl acetate/acetone 1:0 to 4:1 to give a
brown gum. This was further purified by preparative thin layer
chromatography using iso-hexanes/ethyl acetate 1:3 to afford the
title compound (1.8 mg, 3%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta.0.84-1.02 (m, 6H), 1.26 (app s, 3H), 1.67-1.72
(m, 3H), 1.83-1.96 (m, 1H), 2.00-2.08 (m, 1H), 2.63-2.74 (m, 1H),
3.24 (d, J=4.5 Hz, 1H), 3.50 (d, J=7.6 Hz, 1H), 4.21 (app t, J=10.3
Hz, 1H), 4.53-4.61 (m, 1H), 5.55 (app t, J=7.3 Hz, 1H), 6.04-6.11
(m, 2H), 6.32 (d, J=7.8 Hz, 1H), 6.62 (d, J=16.3 Hz, 1H), 6.72-6.80
(m, 1H), 7.29-7.33 (m, 1H), 7.45 (d, J=8.5 Hz, 1H), 7.72 (d, J=8.3
Hz, 1H), 7.97-8.02 (m, 1H). LCMS (m/z) 552.2 [M+H], Tr=1.86
min.
Example 17
##STR00118##
Under argon, a solution of diisopropylamine (2.51 g, 24.8 mmol) in
tetrahydrofuran (150 mL) was cooled in an ice water bath. A
solution of n-butyllithum in hexanes (2.5 M, 9.7 mL, 24 mmol) was
added dropwise over 2 min, and the resulting solution was stirred
for 10 additional min. The solution was then cooled to -78.degree.
C. in a CO.sub.2:acetone bath, and methyl
2,2-dimethyl-3-oxobutanoate (3.2 g, 22 mmol) was added dropwise
over 30 s. The solution was stirred for an additional 15 min, and
N-phenyl-bis(trifluoromethanesulfonimide) (8.4 g, 23.5 mmol) was
added as a solution in tetrahydrofuran (20 mL) via cannula over 5
min, washing with an additional portion of tetrahydrofuran (10 mL).
The resulting solution was stirred for 10 min and was removed from
the cold bath. After stirring an additional 1 h, the reaction
mixture was concentrated in vacuo and diluted with diethyl ether
(150 mL). The organic phase was washed with 1 M aqueous sodium
hydroxide (1.times.100 mL, 1.times.30 mL), dried over anhydrous
magnesium sulfate, filtered, and concentrated to afford the title
compound (6.2 g, 100%) as an amber liquid that was used without
further purification.
##STR00119##
A solution of 17a (6.2 g, 22 mmol) in anhydrous pyridine (11 mL,
140 mmol) was heated to 70.degree. C. After 18.5 h, the temperature
was increased to 90.degree. C. After stirring for a total of 72 h,
the reaction mixture was partitioned between a stirred mixture of
diethyl ether (100 mL) and 3 M aqueous hydrochloric acid (100 mL).
The phases were separated, and the organic layer was washed with
saturated aqueous sodium bicarbonate (75 mL), dried over magnesium
sulfate, filtered, and concentrated to afford the title compound
(2.7 g, 97%) as a slightly brown liquid that was used without
further purification.
##STR00120##
Under argon, bis(cyclopentadienyl)zirconium chloride hydride (290
mg, 1.1 mmol) was cooled in an ice water bath. A solution of 17b
(1.4 g, 11.1 mmol) and pinacolborane (2.4 mL, 16.5 mmol) in
dichloromethane (3 mL) was added by cannula, washing with an
additional portion of dichloromethane (2 mL). The resulting mixture
was removed from the cold bath and was stirred for 72 h at RT. The
reaction was then diluted with ethyl acetate (50 mL), quenched with
dropwise water (5 mL), and was further diluted with water (50 mL).
The organic and aqueous phases were separated, and the aqueous
phase was extracted with ethyl acetate (30 mL). The combined
organic phases were dried over anhydrous sodium sulfate, filtered,
and concentrated to afford a crude residue that was purified by
silica gel chromatography (5 to 15% ethyl acetate in iso-hexanes)
to afford the title compound (1.6 g, 57%) as a colorless oil that
crystallized on standing at -15.degree. C.
##STR00121##
A mixture of 6-bromo-3-chloro-isoquinoline (485 mg, 2.0 mmol),
bis(pinacolato)diboron (560 mg, 2.2 mmol), potassium acetate (392
mg, 4.0 mmol) and
[1,1-bis(diphenylphosphino)ferrocene]dichloropalladium(II), complex
with dichloromethane (82 mg, 0.1 mmol) in 1,4-dioxane (4 mL) was
heated at 160.degree. C. in a microwave for 1 hour. The reaction
mixture was filtered through Celite and the filter pad was washed
with ethyl acetate. The filtrate was evaporated and the residue was
purified by silica gel chromatography using a gradient of
dichloromethane to dichloromethane/methanol 9:1 to afford the title
compound (545 mg, 94%).
##STR00122##
A solution of 17d (1.74 g, 6.0 mmol) in tetrahydrofuran (45 mL) was
stirred at 0.degree. C. under nitrogen. Sodium hydroxide solution
(2 M, 9 mL, 18 mmol) and hydrogen peroxide (30%, 2.5 mL, 24 mmol)
were added dropwise. The reaction mixture was stirred at 0.degree.
C. for 30 min. Water (30 mL) was added and the solution was
acidified to pH 1 with 2 M hydrochloric acid. Sodium metabisulfite
solution (1 M) was added and the mixture was extracted with ethyl
acetate. The organic extracts were combined, washed with water and
brine. The organic solution was filtered through a hydrophobic frit
and the filtrate was evaporated to afford the title compound (662
mg, 61%) as an off-white solid.
##STR00123##
To a solution of 17e (1.25 g, 7.07 mmol) and
tributyl-(1-ethoxy-vinyl)-tin (5.09 g, 4.77 mL, 14.1 mmol) in
1,4-dioxane (15 mL) was added bis(triphenylphosphine)palladium(II)
dichloride (992 mg, 1.41 mmol) and the reaction mixture was heated
at 160.degree. C. in a microwave for 30 min. The reaction mixture
was filtered through Celite and the filter pad was washed with
ethyl acetate. The filtrate was evaporated and the residue was
purified by silica gel chromatography using a gradient of
iso-hexanes/ethyl acetate 9:1 to 3:1 to afford
3-(1-ethoxy-vinyl)-isoquinolin-6-ol (670 mg) as a gum which was
used crude in the next step. 3-(1-Ethoxy-vinyl)-isoquinolin-6-ol
(670 mg) was suspended in 1,4-dioxane (4 mL) and 4 M hydrogen
chloride in 1,4-dioxane (8 mL) was added and the reaction mixture
was stirred at RT for 30 min. The solvent was evaporated to afford
the title compound (584 mg, 44% over 2 steps) as a white solid.
##STR00124##
A solution of 17f (384 mg, 2.05 mmol) and pyridine (0.51 mL, 6.16
mmol) in dichloromethane (30 mL) was stirred in a salt-ice bath for
5 min. Trifluoromethanesulfonic anhydride (0.415 mL, 2.46 mmol) was
added dropwise and the mixture was warmed to RT over 20 min.
Additional trifluoromethanesulfonic anhydride (0.1 mL, 0.6 mmol)
was added and the reaction mixture was stirred at RT for 10 min.
Saturated ammonium chloride solution was added and the mixture was
extracted with dichloromethane. The organic extracts were combined,
dried over sodium sulfate, filtered and evaporated. The residue was
purified by silica gel chromatography using iso-hexanes/ethyl
acetate 19:1 to afford the title compound (300 mg, 44%) as a white
solid.
##STR00125##
To dichloro (p-cymene) ruthenium (II) dimer (2 mg, 0.003 mmol) in
water (2 mL) at RT was added
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (3 mg, 0.008
mmol). The system was degassed for 15 min and then heated to
70.degree. C. for 1.5 h. The reaction was cooled and a solution of
17g (224 mg, 0.70 mmol) in degassed anhydrous tetrahydrofuran (1
mL) was added followed by sodium formate (237 mg, 3.50 mmol). The
system was degassed for 2 min and then heated at 40.degree. C. for
1 hour. After cooling to RT, water was added and the water was
extracted with dichloromethane (2.times.). The combined organic
layers were dried through a hydrophobic frit and concentrated in
vacuo. The product was purified by silica gel chromatography using
iso-hexanes/ethyl acetate 1:1 to afford the title compound (220 mg,
97%) as a brown oil.
##STR00126##
A round bottom flask was charged with 17h (100 mg, 0.31 mmol),
(E)-2,2-dimethyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-but-3-e-
noic acid methyl ester (91 mg, 0.36 mmol),
bis[(dicyclohexyl)(4-dimethylaminophenyl)phosphine]palladium(II)
chloride (13 mg, 0.02 mmol), potassium phosphate tribasic (198 mg,
0.93 mmol) and lithium chloride (40 mg, 0.93 mmol). The system was
flushed with nitrogen and cyclopentyl methyl ether (1 mL) and water
(0.5 mL) were added. The reaction was heated for 1 hour at
90.degree. C. and then cooled to RT. Ethyl acetate was added and
the solution was washed with water. The organic layer was dried
over magnesium sulfate, filtered and concentrated in vacuo. The
residue was purified by silica gel chromatography using
iso-hexanes/ethyl acetate 1:2 to give the title compound (70 mg,
75%) as a yellow oil.
##STR00127##
To 17i (140 mg, 0.47 mmol) in tetrahydrofuran (1 mL), methanol (0.5
mL) and water (0.5 mL) was added lithium hydroxide monohydrate (39
mg, 0.93 mmol) at RT. The reaction was stirred for 3 h and quenched
by adding 2 M aqueous hydrochloric acid (0.5 mL). The reaction was
concentrated in vacuo, followed by co-evaporation from methanol and
then toluene. The ensuing yellow solid was used crude.
##STR00128##
To 1e (250 mg, 0.47 mmol) in anhydrous dichloromethane (5 mL) at
0.degree. C. and under an atmosphere of nitrogen, was added
trimethylsilyl trifluoromethanesulfonate (128 .mu.L, 0.70 mmol).
The reaction mixture was stirred at 0.degree. C. for 2 h before
adding N,N-diisopropylethylamine (252 .mu.L, 1.41 mmol) and then
concentrated in vacuo, and co-evaporated with toluene to afford
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester as a white
solid. To 17j (134 mg, 0.47 mmol) in anhydrous
N,N-dimethylformamide (2 mL) at RT and under an atmosphere of
nitrogen was added N,N-diisopropylethylamine (420 .mu.L, 2.35 mmol)
and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (250 mg, 0.66 mmol). The solution
was stirred at RT for 3 min before adding a solution of
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester in anhydrous
N,N-dimethylformamide (2 mL). The reaction was stirred for 16 h.
The reaction was quenched with water and extracted with ethyl
acetate (2.times.). The combined organic layers were dried over
sodium sulfate, filtered and concentrated in vacuo. The residue was
purified by silica gel chromatography using iso-hexanes/acetone 1:1
to give the title compound (220 mg, 67% over 3 steps) as an off
white solid.
##STR00129##
To 17k (220 mg, 0.31 mmol) in tetrahydrofuran (3 mL), methanol (1.5
mL) and water (1.5 mL) was added lithium hydroxide monohydrate (53
mg, 1.26 mmol) at RT. The reaction was stirred for 1.25 h and
quenched by adding 2 M aqueous hydrochloric acid (0.64 mL). The
reaction was concentrated in vacuo, followed by co-evaporation from
methanol (2.times.) and then acetonitrile (6.times.). The resulting
residue was dissolved in anhydrous N,N-dimethylformamide (4 mL),
and added via syringe pump over 2 h to a suspension of
2-methyl-6-nitrobenzoic anhydride (271 mg, 0.79 mmol),
4-dimethylaminopyridine (288 mg, 2.36 mmol) and powdered 4 .ANG.
molecular sieves (3 g) in 1,2-dichloroethane (103 mL) at 50.degree.
C. Following the addition the reaction was stirred at 50.degree. C.
for 3 h, cooled to RT and filtered through Celite. The filtrate was
concentrated to 1/3 of its volume, diluted with dichloromethane and
washed with water (2.times.). The organic layer was dried through a
hydrophobic frit and concentrated in vacuo. The residue was
purified by silica gel chromatography using iso-hexanes/acetone 1:1
to give an off white solid. The solid was triturated twice with
diethyl ether and vacuum dried for 16 h to afford the title
compound (65 mg, 38% over 2 steps) as a white powder. .sup.1H NMR
(300 MHz, CDCl.sub.3) 0.93 (d, J=6.7 Hz, 3H), 0.99 (d, J=6.7 Hz,
3H), 1.41 (s, 3H), 1.51 (s, 3H), 1.61 (d, J=7.1 Hz, 3H), 1.76 (d,
J=6.7 Hz, 3H), 1.76 (m, 1H), 1.78-2.03 (m, 3H), 2.07-2.17 (m, 1H),
2.62-2.76 (m, 1H), 3.62-3.79 (m, 2H), 4.21-4.31 (m, 1H), 4.52-4.62
(m, 1H), 5.82-6.01 (m, 1H), 6.13 (q, J=6.7 Hz, 1H), 6.27 (d, J=16.1
Hz, 1H), 6.30-6.41 (m, 2H), 6.64 (d, J=16.1 Hz, 1H), 7.53 (d, J=8.5
Hz, 1H), 7.71 (s, 1H), 7.82 (s, 1H), 7.90 (d, J=8.5 Hz, 1H), 9.16
(s, 1H). LCMS (m/z) 549.9 [M+H], Tr=5.21 min.
Example 18
##STR00130##
To Compound 17 (40 mg, 0.073 mmol) in ethyl acetate (10 mL) was
added 10% palladium on carbon (30 mg) at RT. The system was purged
with hydrogen and stirred vigorously for 16 h. The suspension was
filtered through Celite and the filtrate concentrated in vacuo. The
residue was purified by silica gel chromatography using
iso-hexanes/acetone 1:1 to afford Compound 18 (8 mg, 20%) as a
white solid. .sup.1H NMR (300 MHz, CDCl.sub.3) 0.92 (d, J=6.5 Hz,
3H), 1.00 (d, J=6.5 Hz, 3H), 1.27 (s, 3H), 1.31 (s, 3H), 1.48 (d,
J=7.1 Hz, 3H), 1.64-1.79 (m, 1H), 1.75 (d, J=6.7 Hz, 3H), 1.80-2.12
(m, 5H), 2.45 (td, J=12.7, 3.6 Hz, 1H), 2.68-2.93 (m, 2H),
3.62-3.78 (m, 2H), 3.86 (d, J=12.0 Hz, 1H), 4.37 (app t, J=8.3 Hz,
1H), 4.50-4.61 (m, 1H), 5.87-6.00 (m, 1H), 6.18 (q, J=6.7 Hz, 1H),
6.38 (d, J=8.7 Hz, 1H), 6.58 (d, J=8.3 Hz, 1H), 7.36 (d, J=8.5 Hz,
1H), 7.42 (s, 1H), 7.64 (s, 1H), 7.83 (d, J=8.5 Hz, 1H), 9.14 (s,
1H). LCMS (m/z) 552.2 [M+H], Tr=5.11 min.
Example 19
##STR00131##
To a solution of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid (339 mg, 0.920 mmol)
and (R)-1-(3-vinyl-isoquinolin-6-yl)-ethanol (183 mg, 0.920 mmol)
in dichloromethane (4.6 mL) were added
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (211
mg, 1.10 mmol) and 4-dimethylaminopyridine (56.2 mg, 0.46 mmol) at
23.degree. C. under an argon atmosphere. After 21 h, the reaction
mixture was purified directly by silica gel flash column
chromatography to afford the title compound (224 mg, 42%) as a
light tan solid.
##STR00132##
To 19a (800 mg, 1.38 mmol) in anhydrous dichloromethane (12 mL) at
0.degree. C. and under an atmosphere of nitrogen, was added
trimethylsilyl trifluoromethanesulfonate (374 .mu.L, 2.07 mmol).
The reaction mixture was stirred at 0.degree. C. for 2 h before
adding N,N-diisopropylethylamine (480 .mu.L, 2.75 mmol) and then
concentrated in vacuo to afford a white solid. To the solid was
added a solution of (E)-(R)-2-ethyl-pent-3-enoic acid (188 mg, 1.65
mmol) in anhydrous acetonitrile (12 mL) followed by
N,N-diisopropylethylamine (240 .mu.L, 1.38 mmol) and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uranium
hexafluorophosphate methanaminium (733 mg, 1.93 mmol). The reaction
was stirred at RT for 3 h and concentrated in vacuo. The residue
was purified by silica gel chromatography using iso-hexanes/ethyl
acetate 1:2 then 1:5 to give the title compound (250 mg, 74% over 2
steps) as a viscous yellow oil.
##STR00133##
To 19b (200 mg, 0.34 mmol) in anhydrous toluene (113 mL) at
70.degree. C. and under an atmosphere of nitrogen, was added
Hoveyda-Grubbs 2.sup.nd generation catalyst (42 mg, 0.07 mmol). The
reaction was heated at 120.degree. C. for 2 h after which an
additional amount of the Hoveyda-Grubbs 2.sup.nd generation
catalyst (30 mg, 0.05 mmol) was added. Following a further 2 h at
120.degree. C. the reaction was cooled to RT and potassium
isocyanoacetate (83 mg) in methanol (2 mL) was added. The reaction
was stirred for 1 hour, silica added and then concentrated in
vacuo. The residue was purified by silica gel chromatography using
ethyl acetate/acetone 3:1 to afford a brown solid. This was
purified further by reverse phase preparative HPLC to give the
title compound (2.2 mg, 1%) as a white solid. .sup.1H NMR (300 MHz,
CD.sub.3OD) 0.97 (d, J=6.5 Hz, 3H), 1.02 (d, J=6.5 Hz, 3H), 1.11
(t, J=7.4 Hz, 3H), 1.64 (d, J=7.1 Hz, 3H), 1.70 (d, J=6.7 Hz, 3H),
1.71-2.06 (m, 7H), 2.66-2.82 (m, 1H), 3.13-3.22 (m, 1H), 3.75-3.85
(m, 1H), 4.30-4.48 (m, 2H), 5.51-5.67 (m, 1H), 6.07 (q, J=6.3 Hz,
1H), 6.48-6.57 (m, 1H), 6.61 (d, J=16.5 Hz, 1H), 7.47 (s, 1H), 7.58
(d, J=8.7 Hz, 1H), 7.89 (s, 1H), 8.07 (d, J=8.7 Hz, 1H), 9.15 (s,
1H). LCMS (m/z) 550.2 [M+H], Tr=1.67 min.
Example 20
##STR00134##
To 19a (390 mg, 0.67 mmol) in anhydrous dichloromethane (7 mL) at
0.degree. C. and under an atmosphere of nitrogen, was added
trimethylsilyl trifluoromethanesulfonate (182 .mu.L, 1.01 mmol).
The reaction mixture was stirred at 0.degree. C. for 1 hour before
quenching with a saturated aqueous solution of sodium hydrogen
carbonate and extracting with ethyl actate (2.times.). The organic
layers were dried over sodium sulfate, filtered and concentrated in
vacuo to afford a white solid. The solid was dissolved in anhydrous
acetonitrile (4 mL) and 2-methyl-but-3-enoic acid (81 mg, 0.81
mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(184 mg, 0.94 mmol) and 1-hydroxybenzotriazole monohydrate (103 mg,
0.67 mmol) were added. The reaction was stirred at RT for 16 h and
concentrated in vacuo. The residue was purified by silica gel
chromatography using iso-hexanes/ethyl acetate 1:1 then 1:3 to give
the title compound (259 mg, 69%) as a clear viscous oil.
##STR00135##
To 20a (250 mg, 0.44 mmol) in degassed anhydrous toluene (148 mL)
was added 2,6-dichlorobenzoquinone (8 mg, 0.04 mmol). The mixture
was heated to 105.degree. C. and a solution of Hoveyda-Grubbs
2.sup.nd generation catalyst (83 mg, 0.13 mmol) in anhydrous
toluene (20 mL), was added via syringe pump over 2 h. An additional
amount of Hoveyda-Grubbs 2.sup.nd generation catalyst (28 mg) was
added and the reaction heated to reflux for 1 h. The reaction was
cooled to RT, silica gel was added and concentrated in vacuo. The
residue was purified by silica gel chromatography using ethyl
acetate/acetone 1:0 then 3:1 to afford an oil. This was triturated
with diethyl ether and a few drops of ethyl acetate to afford a
brown solid which was further purified by preparative TLC using
ethyl acetate/acetone 5/1 to give the title compound (9 mg, 1%) as
an off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3) 0.97 (d,
J=6.7 Hz, 3H), 1.01 (d, J=6.7 Hz, 3H), 1.38 (d, J=6.7 Hz, 3H), 1.62
(d, J=7.1 Hz, 3H), 1.70 (d, J=6.9 Hz, 3H), 1.72-1.82 (m, 2H),
1.91-2.01 (m, 2H), 2.03-2.12 (m, 1H), 2.62-2.74 (m, 1H), 3.35 (app
t, J=7.4 Hz, 1H), 3.59 (d, J=12.5 Hz, 1H), 3.65-3.77 (m, 1H), 4.26
(app t, J=8.9 Hz, 1H), 4.51-4.60 (m, 1H), 5.68-5.79 (m, 1H), 6.06
(q, J=6.7 Hz, 1H), 6.16 (d, J=9.2 Hz, 1H), 6.27 (d, J=9.2 Hz, 1H),
6.40 (dd, J=16.0, 7.8 Hz, 1H), 6.71 (d, J=16.0 Hz, 1H), 7.39 (dd,
J=8.5, 1.3 Hz, 1H), 7.58 (s, 1H), 7.74 (s, 1H), 7.93 (d, J=8.5 Hz,
1H), 9.16 (s, 1H). LCMS (m/z) 536.2 [M+H], Tr=1.63 min.
Example 21
##STR00136##
21a was prepared in the same manner as 20a replacing
2-methyl-but-3-enoic acid with (R)-2-methyl-pent-4-enoic acid
(prepared as described in Synlett 2002, No 12, 2039-2040, 82 mg,
0.72 mmol), to afford the title compound (280 mg, 67%) as a white
foam.
##STR00137##
To 21a (250 mg, 0.43 mmol) in anhydrous toluene (144 mL) at RT and
under an atmosphere of nitrogen, was added Hoveyda-Grubbs 2.sup.nd
generation catalyst (54 mg, 0.09 mmol). The reaction was heated at
120.degree. C. for 2 h after which an additional amount of the
Hoveyda-Grubbs 2.sup.nd generation catalyst (25 mg, 0.04 mmol) was
added. Following a further 1 h at 120.degree. C. the reaction was
cooled to RT and concentrated in vacuo. The residue was purified by
silica gel chromatography using ethyl acetate/acetone 1:0 then 3:1
to afford a brown solid. This was triturated with diethyl
ether/ethyl acetate 3:1 to give the title compound (50 mg, 22%) as
an off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3) 0.91 (d,
J=6.7 Hz, 3H), 0.99 (d, J=6.7 Hz, 3H), 1.29 (d, J=7.1 Hz, 3H), 1.59
(d, J=6.7 Hz, 3H), 1.67 (d, J=6.5 Hz, 3H), 1.70-1.84 (m, 1H),
1.92-2.15 (m, 4H), 2.33-2.48 (m, 1H), 2.63-2.98 (m, 3H), 3.53 (d,
J=12.2 Hz, 1H), 3.60-3.72 (m, 1H), 4.42 (dd, J=6.7, 2.2 Hz, 1H),
4.52-4.61 (m, 1H), 5.62-5.74 (m, 1H), 6.11 (q, J=6.7 Hz, 1H), 6.16
(q, J=8.9 Hz, 1H), 6.42 (d, J=8.5 Hz, 1H), 6.53-6.66 (m, 1H), 6.75
(d, J=16.1 Hz, 1H), 7.37 (dd, J=8.7, 1.0 Hz, 1H), 7.80-7.96 (m,
3H), 9.16 (s, 1H). LCMS (m/z) 550.3 [M+H], Tr=1.50 min.
Example 22
##STR00138##
To a stirred slurry of 7-bromo-2-chloro-quinoline (8.10 g, 33.4
mmol) and sodium iodide (50.0 g, 334 mmol) in acetonitrile (27 mL)
was slowly added acetyl chloride (3.56 mL, 50.0 mmol). The flask
was stoppered and sealed and heated at 80.degree. C. for 3 h before
being allowed to cool. The mixture was treated sequentially with
10% w/w aqueous potassium carbonate solution (80 mL), 5% w/w
aqueous sodium sulfite solution (80 mL) and saturated aqueous
sodium thiosulfate solution (80 mL) and the mixture extracted with
dichloromethane (2.times.). The combined organic extracts were
dried over sodium sulfate, filtered and evaporated to give a crude
7-bromo-2-iodo-quinoline. To the quinoline was added
tributyl(1-ethoxyvinyl)tin (13.6 mL, 40.1 mmol), 1,4-dioxane (67
mL) and bis(triphenylphosphine)palladium(II) dichloride (2.37 g,
3.34 mmol) and the reaction mixture heated at 100.degree. C. for 5
h before allowing to cool. 2 M Aqueous hydrochloric acid (67 mL)
was added and the reaction stirred for 1 h. The mixture was
filtered and the solids washed with ethyl acetate and the filtrate
evaporated to remove organics. The residue was extracted with ethyl
acetate (3.times.) and the combined organic extracts were dried
over sodium sulfate, filtered and evaporated. The product was
purified on silica gel doped with 10% w/w potassium carbonate
eluting with a gradient of 0 to 6% ethyl acetate in iso-hexanes to
afford the title compound (5.5 g, 66%) as a white solid.
##STR00139##
Dichloro (p-cymene) ruthenium(II) dimer (61 mg, 0.100 mmol) and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (88 mg, 0.012
mmol) was suspended in degassed water (40 mL) and the mixture was
degassed with nitrogen for 5 min. The mixture was stirred at
70.degree. C. under nitrogen for 90 min. The resulting turbid
orange mixture was allowed to cool to RT. 22a (5.00 g, 20 mmol) in
degassed tetrahydrofuran (40 mL) was added followed by sodium
formate (6.8 g, 100 mmol) and the reaction mixture was degassed
with nitrogen for 5 min. The reaction mixture was vigorously
stirred at 40.degree. C. for 4 h and allowed to cool. It was then
diluted with ethyl acetate and water and the organic layer was
separated, washed with water and brine, dried over sodium sulfate,
filtered and evaporated. The residue was purified by silica gel
chromatography using a gradient of 0% to 30% ethyl acetate in
iso-hexanes to afford the title compound (4.96 g, 98%) as an
off-white solid.
##STR00140##
To a solution of 22b (1.00 g, 3.97 mmol) and triethylamine (1.65
mL, 11.9 mmol) in anhydrous dichloromethane at 0.degree. C., was
added acetic anhydride (0.75 mL, 7.93 mmol) and
4-(dimethylamino)pyridine (24 mg, 0.197 mmol). The reaction mixture
was stirred and allowed to warm to RT. After 1.5 h water (100 mL)
was added and the layers separated. The aqueous phase was
re-extracted with dichloromethane (2.times.100 mL) and the combined
organics were washed with brine (100 mL), dried over magnesium
sulfate, filtered and concentrated in vacuo to give a crude
residue. This was purified by silica gel chromatography using
iso-hexanes (66 mL) then iso-hexanes/ethyl acetate 95:5 (300 mL),
then iso-hexanes/ethyl acetate 9:1 (1066 mL) to yield the title
compound (1.16 g, 99%) as a colorless oil.
##STR00141##
To a solution of acetic acid 22c (1.16 g, 3.95 mmol) in anhydrous
acetonitrile was added palladium(II) acetate (89 mg, 0.395 mmol),
2,2-dimethyl-but-3-enoic acid (496 mg, 4.35 mmol),
tri(o-tolyl)phosphine (241 mg, 0.790 mmol) and triethylamine (1.09
mL, 7.90 mmol) then the mixture was heated in the microwave at
100.degree. C. for 20 min. The reaction mixture was concentrated in
vacuo then water (200 mL) was added and the organics extracted with
ethyl acetate (2.times.50 mL). The combined organics were dried
over sodium sulfate, filtered and concentrated in vacuo to give a
crude residue. This was purified by silica gel chromatography using
iso-hexanes/ethyl acetate 95:5 (300 mL), then iso-hexanes/ethyl
acetate 9:1 (1066 mL) then iso-hexanes/ethyl acetate 3:1 (1066 mL)
to yield the title compound (792 mg, 61%) as a white solid.
##STR00142##
A solution of 1e (1.29 g, 2.42 mmol) in dichloromethane (12 mL) was
cooled in an ice-water bath under nitrogen. Trimethylsilyl
trifluoromethanesulfonate (0.715 mL, 4.84 mmol) was added dropwise,
and the resulting solution was stirred for 1 h. The reaction was
quenched with N,N-diisopropylethylamine (1.69 mL, 9.68 mmol) and
the reaction mixture was concentrated in vacuo to give
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester as a white
solid which was used without further purification. To a solution of
22d (792 mg, 2.42 mmol) in N,N-dimethylformamide (10 mL) at
0.degree. C. under nitrogen was added N,N-diisopropylethylamine
(2.11 mL, 12.1 mmol) and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (1.01 g, 2.66 mmol). The
resulting mixture was stirred at RT for 15 min then re-cooled to
0.degree. C. and
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester generated in
the previous step, was added as a solution in N,N-dimethylformamide
(10.7 mL). The reaction mixture was then allowed to warm to RT with
stirring. After 1 h the mixture was diluted with ethyl acetate (100
mL) and water (100 mL). The phases were separated and the aqueous
was extracted with ethyl acetate (100 mL). The combined organics
were washed with brine (100 mL), dried over magnesium sulfate,
filtered and concentrated in vacuo to give a crude residue. This
was purified by silica gel chromatography using iso-hexanes/acetone
95:5 (1000 mL) then iso-hexanes/acetone 9:1 (1 L) then
iso-hexanes/acetone 85:15 (1 L) then iso-hexanes/acetone 3:1 (1 L)
then iso-hexanes/acetone 7:3 (1 L) to yield the title compound
(1.11 g, 62%) as a colorless solid.
##STR00143##
A solution of 22e (1.11 g, 1.50 mmol) in tetrahydrofuran (14.7 mL)
was cooled in an ice-water bath and methanol (7.4 mL), water (7.4
mL), and lithium hydroxide monohydrate (252 mg, 6.0 mmol) were then
added. The mixture was stirred for 1 h in the ice-water bath and
then quenched with aqueous 1 M hydrochloric acid (6 mL, 6.0 mmol).
The resulting solution was concentrated in vacuo, and the crude
product was concentrated from methanol (4.times.250 mL) then
acetonitrile (4.times.250 mL) and toluene (5.times.250 mL). The
solid isolated was then left on the freeze dryer overnight to
afford the title compound (853 mg, quantitative yield) as a white
solid.
##STR00144##
Under nitrogen, 2-methyl-6-nitrobenzoic anhydride (2.59 g, 7.51
mmol) and 4-(dimethylamino)pyridine (1.38 g, 11.3 mmol) were
dissolved in 1,2-dichloroethane (500 mL) and the resulting solution
was heated to 50.degree. C. The crude seco-acid from the previous
step, 22f (853 mg, 1.50 mmol) was dissolved in
N,N-dimethylformamide (19 mL) and added to the reaction mixture
dropwise via syringe pump over 6 h. After the addition was
complete, the syringe pump was then rinsed with additional
N,N-dimethylformamide (3 mL) and the reaction mixture was stirred
at 50.degree. C. for 40 min. After this time the mixture was
concentrated to give a residue. This was purified by silica gel
chromatography using iso-hexanes (66 mL), then iso-hexanes/acetone
4:1 (726 mL), then iso-hexanes/acetone 7:3 (726 mL), then
iso-hexanes/acetone 3:2 (726 mL). This initial column gave impure
product (1.10 g) which contained N,N-dimethylformamide. To this was
added brine (200 mL) and the organics extracted with ethyl acetate
(2.times.100 mL). The combined organics were dried over magnesium
sulfate, filtered and concentrated in vacuo to give impure product
(900 mg) which does not contain N,N-dimethylformamide. This residue
was purified by a second silica gel chromatography using
iso-hexanes/acetone 95:5 (6 L), then iso-hexanes/acetone 93:7 (1.5
L), then iso-hexanes/acetone 88:12 (3 L), then iso-hexanes/acetone
82:16 until all the product eluted from the column. Two batches of
the desired product in 87% purity (batch A) and 77% purity (batch
B) were isolated. Batch A was triturated twice with 100% diethyl
ether to give the title compound (241 mg) in approximately 90-95%
purity. Purification of Batch B by preparative reverse phase HPLC
gave the title compound (181 mg) in approximately >95% purity.
Batch A and B were combined to give the title compound (422 mg,
51%) as a white solid. .sup.1H NMR (500 MHz, CD.sub.3OD) 0.94 (app
t, J=6.7 Hz, 6H), 1.34 (s, 3H), 1.48 (s, 3H), 1.58 (d, J=7.3 Hz,
3H), 1.62-1.75 (m, 5H), 1.82-1.91 (m, 1H), 1.92-2.03 (m, 2H),
2.66-2.78 (m, 1H), 3.78 (app d, J=8.5 Hz, 1H), 4.25-4.33 (m, 1H),
4.34-4.43 (m, 1H), 5.71 (q, J=7.3 Hz, 1H), 5.89 (q, J=6.7 Hz, 1H),
6.24 (d, J=16.0 Hz, 1H), 6.46 (d, J=16.0 Hz, 1H), 7.27 (br d, J=9.8
Hz, 1H), 7.38 (d, J=8.6 Hz, 1H), 7.60 (s, 1H), 7.75 (ABq,
.DELTA..delta..sub.AB=0.03, J.sub.AB=8.54 Hz, 2H), 8.18 (d, J=7.9
Hz, 1H). LCMS (m/z) 550.1 [M+H], Tr=2.24 min.
Example 23
##STR00145##
A solution of 6-bromo-3-chloro-isoquinoline (2.0 g, 8.25 mmol) in
n-propanol (90 mL) was prepared and potassium vinyltrifluoroborate
(1.11 g, 8.25 mmol) was added. The solution was purged with
nitrogen for 10 min before addition of
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II),
complex with dichloromethane (128 mg, 0.157 mmol) and triethylamine
(1.15 mL, 8.25 mmol). The reaction mixture was then purged with
nitrogen for a further 3 min before heating to reflux for 1 h. The
reaction mixture was then allowed to cool to RT and water was
added. The organics were then extracted with diethyl ether
(3.times.150 mL). The combined organics were dried over magnesium
sulfate, filtered and concentrated in vacuo to give a crude
residue. This was purified by silica gel chromatography, using
iso-hexanes/diethyl ether 9:1 to give the title compound (1.26 g,
80%) as an oil.
##STR00146##
A suspension of 23a (1.24 g, 6.54 mmol) in methanol (125 mL) and
dichloromethane (125 mL) was cooled to -78.degree. C. The reaction
was ozonised until a blue colour persisted (in approximately 15
min), then nitrogen was bubbled through the reaction mixture for 15
min to purge the ozone. The reaction was then treated with solid
sodium bicarbonate (549 mg, 6.54 mmol) and dimethyl sulfide (1.31
mL, 1.31 mmol). The mixture was allowed to warm to RT and after 3 h
the reaction mixture was concentrated in vacuo. Water (200 mL) was
added to the residue and the aqueous layer extracted with
dichloromethane (3.times.200 mL). The combined organics were dried
over magnesium sulfate, filtered and concentrated in vacuo to give
the title compound (1.02 g, 81%) as an oil.
##STR00147##
To a solution of 23b (1.02 g, 5.32 mmol) in tetrahydrofuran (10 mL)
at 0.degree. C. was added trimethyl(trifluoromethyl)silane solution
(3.19 mL, 2 M in tetrahydrofuran, 6.38 mmol) followed by
tetrabutylammonium fluoride solution (0.053 mL, 1 M in
tetrahydrofuran, 0.053 mmol) and the resulting solution was stirred
at 0.degree. C. for 1.5 h. The reaction mixture was then allowed to
warm to RT and 2 M hydrochloric acid (10 mL) was added and the
mixture stirred for 10 min before addition of water (200 mL). The
aqueous layer was then extracted with ethyl acetate (3.times.100
mL). The organics were combined, dried over sodium sulfate,
filtered and concentrated to give the title compound (1.30 g, 93%)
as a solid.
##STR00148##
To a solution of 23c (1.30 g, 4.97 mmol) in dichloromethane (14.7
mL) at 0.degree. C. was added Dess-Martin periodinane solution
(14.7 mL, 15% in dichloromethane, 6.96 mmol) and the reaction
mixture was stirred and allowed to warm to RT. 1 M Aqueous sodium
metabisulfite (100 mL) was added and the reaction mixture was
stirred for 15 min. To this mixture was added saturated aqueous
sodium bicarbonate solution (100 mL) and the aqueous layer was
extracted with dichloromethane (3.times.100 mL). The combined
organics were washed with saturated aqueous sodium bicarbonate
solution (200 mL) followed by brine (100 mL), dried over magnesium
sulfate, filtered and concentrated in vacuo to give the title
compound (1.0 g, 78%) as a solid.
##STR00149##
A mixture of 23d (686 mg, 2.64 mmol),
(R)-2-methyl-2-propanesulfinamide (400 mg, 3.30 mmol) and
titanium(IV) isopropoxide (1.95 mL, 6.60 mmol) in tetrahydrofuran
(27.4 mL) was heated at reflux for 16 h then allowed to cool to RT.
The reaction mixture was concentrated in vacuo and re-dissolved in
diethyl ether (27.4 mL) then cooled to -78.degree. C. To this
mixture was added L-Selectride.RTM. solution (7.93 mL, 1.0 M in
tetrahydrofuran, 7.93 mmol) and the reaction mixture stirred at
-78.degree. C. for 1 h. Brine (30 mL) was then added and the
mixture allowed to warm to RT. More brine (200 mL) and ethyl
acetate (300 mL) were added and the layers separated. The aqueous
phase was re-extracted with dichloromethane (500 mL). The combined
organics were dried over sodium sulfate, filtered and concentrated
in vacuo to give a crude residue. This was purified by silica gel
chromatography, using gradient elution of dichloromethane to
dichloromethane/methanol 99:1 to give the title compound (553 mg,
57% yield, 94% d.e.).
##STR00150##
To a solution of 23e (553 mg, 1.52 mmol) in methanol (10 mL) was
added 4 M hydrochloric acid solution in 1,4-dioxane (1.52 mL, 6.06
mmol) and the reaction mixture was stirred at RT for 30 min.
Additional 4 M hydrochloric acid solution in 1,4-dioxane (1.52 mL,
6.06 mmol) was added and the reaction mixture stirred at RT for 30
min then concentrated in vacuo to give the title compound (507 mg,
quantitative yield).
##STR00151##
A mixture of 23f (507 mg, 1.52 mmol) and triethylamine (0.84 mL,
6.08 mmol) in dichloromethane (20 mL) was stirred at 0.degree. C.
to give a solution. Then a solution of di-tert-butyl dicarbonate
(497 mg, 2.28 mmol) in dichloromethane (4 mL) was added and the
reaction mixture was stirred and allowed to warm to RT over 24 h.
Water (200 mL) was added and the aqueous layer extracted with
dichloromethane (3.times.100 mL), dried over sodium sulfate,
filtered and concentrated in vacuo to give a residue (816 mg). This
residue was dissolved in dichloromethane (20 mL) and cooled to
0.degree. C. then triethylamine (0.84 mL, 6.08 mmol) was added
followed by a solution of di-tert-butyl dicarbonate (497 mg, 2.28
mmol) in dichloromethane (4 mL) and the reaction mixture was
stirred and allowed to warm to RT over 72 h. Additional
di-tert-butyl dicarbonate (162 mg, 0.74 mmol) was added and
stirring continued for 30 min then water (100 mL) was added and the
aqueous layer extracted with dichloromethane (2.times.50 mL). The
combined organics were dried over sodium sulfate, filtered and
concentrated in vacuo to give a crude residue. This was purified by
silica gel chromatography, using gradient elution of
iso-hexanes/ethyl acetate 9:1 to iso-hexanes/ethyl acetate 1:1 to
give the title compound (140 mg, 25%) as an oil.
##STR00152##
To a solution of 23g (135 mg, 0.37 mmol) in n-propanol (6 mL) was
added potassium vinyltrifluoroborate (55 mg, 0.412 mmol),
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II),
complex with dichloromethane (5.8 mg, 0.007 mmol) and triethylamine
(0.052 mL, 0.37 mmol). The suspension was evacuated and purged with
nitrogen 3 times before heating to 100.degree. C. for 2 h. After
this time additional potassium vinyltrifluoroborate (100 mg, 0.748
mmol) was added and heating continued at 100.degree. C. for 16 h.
After this time, additional [1,1'-bis(diphenylphosphino)ferrocene]
dichloropalladium(II), complex with dichloromethane (9 mg, 0.011
mmol) was added and the reaction mixture was heated in a microwave
reactor at 150.degree. C. for 90 min. After this time additional
[1,1-bis(diphenylphosphino)ferrocene]dichloropalladium(II), complex
with dichloromethane (9 mg, 0.011 mmol) was added and the reaction
mixture was heated in a microwave reactor at 150.degree. C. for 10
min. Water (100 mL) was added to the reaction mixture and the
aqueous layer was then extracted with diethyl ether (2.times.100
mL). The combined organics were dried over sodium sulfate, filtered
and concentrated in vacuo to give the title compound (132 mg,
quantitative yield).
##STR00153##
To a solution of 23h (132 mg, 0.37 mmol) in methanol (2 mL) was
added 4 M hydrochloric acid in 1,4-dioxane (0.37 mL, 0.75 mmol) and
the mixture stirred at RT for 30 min. Additional 4 M hydrochloric
acid in 1,4-dioxane (3.0 mL, 6.08 mmol) was added and stirring
continued for 2 h then the reaction mixture was concentrated in
vacuo to give a residue. The residue was then concentrated from
diethyl ether (2 mL), followed by acetonitrile (10 mL) and this was
repeated twice to give
(S)-2,2,2-trifluoro-1-(3-vinyl-isoquinolin-6-yl)-ethylamine
hydrochloric acid salt (108 mg, quantitative yield). To a
suspension of
(S)-2,2,2-trifluoro-1-(3-vinyl-isoquinolin-6-yl)-ethylamine
hydrochloric acid salt (108 mg, 0.374 mmol) in acetonitrile (2 mL)
at 0.degree. C. under nitrogen was added
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid (138 mg, 0.37 mmol),
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (100
mg, 0.52 mmol) and 1-hydroxybenzotriazole hydrate (63 mg, 20 wt %
in water, 0.37 mmol). The resulting suspension was stirred at
0.degree. C. for 15 min before removing from the ice-water bath and
stirring at RT for 16 h. N,N-Diisopropylethylamine (0.5 mL, 2.87
mmol) was added and the solution was stirred at RT for 2.5 h where
additional
(S)-1-[(S)-2-((S)-2-but-3-enoylamino-3-methyl-butyrylamino)-propionyl]-he-
xahydro-pyridazine-3-carboxylic acid (69 mg, 0.19 mmol) was added
and stirring continued. After 1 h,
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (134
mg, 0.70 mmol) was added and the reaction mixture was stirred at RT
for 1 hour. After this time, the mixture was concentrated in vacuo
and the residue diluted with ethyl acetate (60 mL). Water (50 mL)
was added and the layers separated. The aqueous phase was
re-extracted with ethyl acetate (2.times.10 mL) and the combined
organics were then washed with ammonium chloride (2.times.50 mL),
dried over sodium sulfate, filtered and concentrated in vacuo to
give a crude residue. This was purified by silica gel
chromatography, using gradient elution of iso-hexanes/ethyl acetate
9:1 to iso-hexanes/ethyl acetate 4:1 to iso-hexanes/ethyl acetate
1:1 and finally iso-hexanes/ethyl acetate 0:1. This gave the title
compound (124 mg, 55%).
##STR00154##
To a solution of 23i (124 mg, 0.21 mmol) in toluene (69 mL) was
added Hoveyda-Grubbs 2.sup.nd generation catalyst (13 mg, 0.02
mmol) and the reaction mixture heated at 115.degree. C. for 1.5 h.
Additional Hoveyda-Grubbs 2.sup.nd generation catalyst (13 mg, 0.02
mmol) was added and heating continued at 120.degree. C. for 50 min.
Additional Hoveyda-Grubbs 2.sup.nd generation catalyst (13 mg, 0.02
mmol) was added and heating continued at 120.degree. C. for 30 min.
After this time the mixture was allowed to cool to RT and
concentrated in vacuo. The residue was purified by silica gel
chromatography using a gradient of ethyl acetate to ethyl
acetate/acetone 9:1. Impure product (28.3 mg) was collected which
was further purified by preparative TLC eluting with ethyl
acetate/acetone 95:5 to afford the title compound (4 mg, 3%) as a
white solid. .sup.1H NMR (300 MHz, CD.sub.3OD) 0.99 (d, J=6.7 Hz,
3H), 1.01 (d, J=6.7 Hz, 3H), 1.29-1.39 (m, 1H), 1.69 (d, J=7.4 Hz,
3H), 1.74-2.01 (m, 5H), 2.73 (td, J=12.9, 2.9 Hz, 1H), 2.99-3.11
(m, 1H), 3.74-3.83 (m, 1H), 4.26 (d, J=10.0 Hz, 1H), 4.41 (br d,
J=13.2 Hz, 1H), 5.48 (q, J=7.4 Hz, 1H), 5.84 (q, J=8.3 Hz, 1H),
6.58 (d, J=16.3 Hz, 1H), 6.62-6.71 (m, 1H), 7.47 (s, 1H), 7.76 (d,
J=8.6 Hz, 1H), 7.89 (s, 1H), 8.13 (d, J=8.6 Hz, 1H), 9.22 (s, 1H).
LCMS (m/z) 575.2 [M+H], Tr=1.53 min.
Example 24
##STR00155##
To a solution of (R)-1-(7-bromo-quinolin-2-yl)-ethanol (295 mg,
1.17 mmol) in anhydrous acetonitrile (12 mL) were added but-3-enoic
acid tert-butyl ester (0.44 mL, 2.75 mmol),
bis(tri-tert-butylphosphine)palladium(0) (25 mg, 0.049 mmol) and
N,N-dicyclohexylmethylamine (0.39 mL, 1.83 mmol) then the mixture
was heated at reflux for 2 h. The reaction mixture was concentrated
in vacuo to give a crude residue. This was purified by silica gel
chromatography using iso-hexanes/ethyl acetate 95:5 to yield the
title compound (348 mg, 95%) as a white solid.
##STR00156##
To 24a (363 mg, 1.16 mmol) and
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid (505 mg, 1.26 mmol) in
dichloromethane (20 mL) was added
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (311
mg, 1.62 mmol) and 4-dimethylaminopyridine (142 mg, 1.16 mmol). The
reaction mixture was then stirred at RT for 16 h. To the mixture
was added saturated aqueous ammonium chloride solution (100 mL) and
the aqueous layer extracted with dichloromethane (2.times.50 mL).
The aqueous phase was further washed with dichloromethane (20 mL)
and the combined organics washed with brine (50 mL), dried over
magnesium sulfate, filtered and concentrated in vacuo to give a
crude residue. This was purified by silica gel chromatography using
iso-hexanes/ethyl acetate 4:1 (120 mL) then iso-hexanes/ethyl
acetate 7:3 (855 mL) then iso-hexanes/ethyl acetate 3:2 (540 mL) to
yield the title compound (357 mg, 44%) as a white solid.
##STR00157##
To a solution of 24b (342 mg, 0.492 mmol) in dichloromethane (2.5
mL) at 0.degree. C. was added trimethylsilyl
trifluoromethanesulfonate (0.18 mL, 0.96 mmol) and the pale yellow
solution stirred at 0.degree. C. for 2.75 h. Additional
trimethylsilyl trifluoromethanesulfonate (0.073 mL, 0.45 mmol) was
added and stirring continued for 1.33 h. N,N-Diisopropylethylamine
(0.6 mL, 3.44 mmol) was added and the mixture stirred for 10 min
then concentrated in vacuo to give intermediate
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid
(R)-1-[7-((E)-3-carboxy-propenyl)-quinolin-2-yl]-ethyl ester (266
mg, 0.45 mmol). To a solution of
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid
(R)-1-[7-((E)-3-carboxy-propenyl)-quinolin-2-yl]-ethyl ester (266
mg, 0.45 mmol) in acetonitrile (50 mL) at 0.degree. C. under
nitrogen was added N,N-diisopropylethylamine (0.343 mL, 1.971 mmol)
and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (262 mg, 0.70 mmol). The mixture
was stirred at 0.degree. C. and allowed to warm to RT over 16 h. To
the reaction mixture was added 2 M aqueous hydrochloric acid
solution (20 mL) and the mixture concentrated in vacuo. The aqueous
layer was extracted with dichloromethane/methanol 9:1 (2.times.100
mL). The combined organics were washed with saturated sodium
hydrogen carbonate solution (2.times.200 mL), then brine
(1.times.100 mL), dried over sodium sulfate, filtered and
concentrated in vacuo to give a crude residue. This was purified by
silica gel chromatography using gradient elution of ethyl
acetate/acetone 97:3 to ethyl acetate/acetone 94:6. Collected 45 mg
of impure product which was triturated with diethyl ether to give
the title compound (25.4 mg, 10%) as a pale yellow solid. .sup.1H
NMR (300 MHz, CD.sub.3OD) 0.99 (d, J=6.48 Hz, 3H), 1.01 (d, J=6.7
Hz, 3H), 1.64 (d, J=7.1 Hz, 3H), 1.66-1.72 (m, 1H), 1.74 (d, J=6.9
Hz, 3H), 1.85-2.08 (m, 4H), 2.75 (dd, J=2.9, 12.9 Hz, 1H),
2.97-3.07 (m, 1H), 3.35-3.40 (m, 1H), 3.79-3.88 (m, 1H), 4.23 (d,
J=10.5 Hz, 1H), 4.43 (br d, J=11.2 Hz, 1H), 5.72 (q, J=7.1 Hz, 1H),
5.94 (q, J=6.9 Hz, 1H), 6.41 (d, J=16.7 Hz, 1H), 6.55 (dt, J=4.7,
16.7 Hz, 1H), 7.40 (d, J=8.5 Hz, 1H), 7.65 (s, 1H), 7.75 (dd,
J=1.3, 8.7 Hz, 1H), 7.81 (d, J=8.7 Hz, 1H), 8.21 (d, J=8.5 Hz, 1H).
LCMS (m/z) 522.2 [M+H], Tr=1.90 min.
Example 25
##STR00158##
A cooled (-20.degree. C.) solution of (S)-6-bromo-indan-1-ol
(1.0462 g, 4.910 mmol, prepared as described in WO 2009/003719) in
anhydrous tetrahydrofuran (20 mL) was subsequently treated with
triethylamine (2.7 mL, 19.639 mmol) and methanesulfonyl chloride
(760 .mu.L, 9.820 mmol). After stirring at -30.degree. C. for 2.5
h, a solution of methylamine (2 M in tetrahydrofuran, 25 mL, 50
mmol) was added. After stirring at RT for 22.5 h the reaction
mixture was filtered. The white solid was rinsed with diethyl
ether. The filtrate was evaporated to dryness to afford the title
compound (1.11 g, quantitative yield) as a white solid.
##STR00159##
A solution of 25a (1.11 g, 4.910 mmol) was subsequently treated
with di-tert-butyl dicarbonate (1.714 g, 7.856 mmol) and
triethylamine (690 .mu.L, 4.91 mmol). After stirring for 25.5 h,
the volatiles were removed in vacuo and the residue was purified by
silica gel chromatography using a 50 g Isolute cartridge eluted
with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to 9:1
to afford the title compound (1.4224 g, 89%) as a white solid as a
mixture of rotamers.
##STR00160##
A solution of 25b (1.4224 g, 4.363 mmol),
[1,1-bis(diphenylphosphino)ferrocene]dichloropalladium
dichloromethane complex (71.3 mg, 0.087 mmol), potassium
vinyltrifluoroborate (701.3 mg, 5.236 mmol) and triethylamine (610
.mu.L, 4.363 mmol) in anhydrous n-propanol (40 mL) was degassed by
bubbling nitrogen through for 30 min. The red suspension was then
refluxed for 17.5 h. After cooling to RT, the mixture was quenched
with water and the aqueous layer was extracted with diethyl ether
(2.times.). The organics were combined, dried over sodium sulfate,
filtered and the volatiles were removed in vacuo. The residue was
purified by silica gel chromatography using a 50 g Isolute
cartridge eluted with a continuous gradient of iso-hexanes/diethyl
ether 1:0 to 4:1 to afford the title compound (977.3 mg, 82%) as a
white solid as a mixture of rotamers.
##STR00161##
A cooled (0.degree. C.) solution of 25c (977.3 mg, 3.575 mmol) in
dichloromethane (20 mL) was treated with trifluoroacetic acid (5
mL). After stirring for 30 min at RT the volatiles were removed in
vacuo and the residual trifluoroacetic acid was azeotroped off with
toluene (3.times.) to afford the corresponding ammonium salt as a
pink solid. A cooled (0.degree. C.) solution of the crude ammonium
salt, crude
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid (3.932 mmol) and
N,N-diisopropylethylamine (2.5 mL, 14.300 mmol) in acetonitrile (60
mL) was treated with
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (1.903 g, 5.005 mmol). After
stirring for 20 h at RT, the reaction was quenched with
hydrochloric acid (1 M, 100 mL). The aqueous layer was extracted
with ethyl acetate (2.times.). The organics were combined, dried
over sodium sulfate, filtered and the volatiles were removed in
vacuo. The residue was purified by silica gel chromatography using
a 100 g Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 1:2 to afford the title compound
in a mixture which was dissolved in ethyl acetate and washed with
aqueous potassium carbonate. The organics were dried over sodium
sulfate, filtered and the volatiles were removed in vacuo to afford
the title compound (798.9 mg, 36%) as a white solid as a mixture of
rotamers.
##STR00162##
Compound 25e was prepared in the same manner as 22e using 25d
instead of 1e in 27% yield as a complex mixture of rotamers.
##STR00163##
A solution of 25e (238.3 mg, 0.390 mmol) in dichloroethane (100 mL)
was treated with Hoveyda-Grubbs 2.sup.nd generation catalyst (48.9
mg, 0.078 mmol). After stirring at reflux for 1.5 h, the reaction
was cooled to RT and the volatiles were removed in vacuo. The
residue was purified by silica gel chromatography using a 10 g
Isolute cartridge eluted by gravity with a continuous gradient of
ethyl acetate/methanol 1:0 to 95:5 followed by preparative TLC
eluted with ethyl acetate/methanol 97:3 (2 elutions) to provide the
final compound (10.2 mg, 5%) as a white solid. .sup.1H NMR (300
MHz, CD.sub.3CN) .delta.0.85-0.97 (m, 6H), 1.21-1.35 (m, 8H),
1.42-1.62 (m, 3H), 1.64-1.77 (m, 2H), 1.80-1.89 (m, 2H), 2.27-2.38
(m, 2H), 2.52 (dd, J=7.6, 3.1 Hz, 1H), 2.76 (s, 3H), 2.82-3.06 (m,
2H), 3.27-3.35 (m, 1H), 3.47 (s, 3H), 3.80-3.91 (m, 1H), 4.06 (dd,
J=9.1, 8.2 Hz, 1H), 4.23 (d, J=11.8 Hz, 1H), 4.41 (dd, J=12.9, 3.6
Hz, 1H), 5.44 (app pentet, J=8.0 Hz, 1H), 6.10-6.30 (m, 2H), 6.43
(d, J=16.0 Hz, 1H), 6.88 (d, J=8.0 Hz, 1H), 6.96 (d, J=9.4 Hz, 1H),
7.04 (s, 1H), 7.13-7.28 (m, 2H). LCMS (m/z) 582.3 [M+H], Tr=2.57
min.
Example 26
##STR00164##
A solution of
(S)-4-benzyl-3-((R)-2-methyl-pent-4-enoyl)-oxazolidin-2-one (1.65
g, 6.04 mmol, prepared as in Synlett 2002, 12, 2039-2040) in
ethanol/water (22 mL, 10:1) was treated with rhodium(III) chloride
hydrate (31.6 mg, 0.15 mmol). After stirring at 85.degree. C. for
24 h, the reaction mixture was cooled to RT and the volatiles were
removed in vacuo. The residue was purified by silica gel
chromatography using a 50 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/diethyl ether 1:0 to 4:1 to
afford the title compound (892.7 mg, 54%) as a colorless oil.
##STR00165##
A cooled (0.degree. C.) solution of 26a (893 mg, 3.27 mmol) in
tetrahydrofuran/water (30 mL, 2:1) was subsequently treated with
hydrogen peroxide (1.7 mL, 16.33 mmol, 30% in water) and lithium
hydroxide hydrate (274 mg, 6.53 mmol). After stirring for 1.5 h at
0.degree. C., the reaction was quenched with sodium metabisulfite
(6.2 g, 32.66 mmol). After stirring at RT for 40 min, the mixture
was acidified with hydrochloric acid (2 M) and the aqueous layer
was extracted with dichloromethane (2.times.). The organics were
combined, filtered through a phase separator and the volatiles were
removed in vacuo. The residue was purified by silica gel
chromatography using a 25 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to
afford the title compound (307.1 mg, 82%) as a colorless oil.
##STR00166##
Compound 26c was prepared in the same manner as
methyl-((R)-6-vinyl-indan-1-yl)-carbamic acid tert-butyl ester
using (R)-1-(7-bromo-quinolin-2-yl)-ethanol instead of
((R)-6-bromo-indan-1-yl)-methyl-carbamic acid tert-butyl ester in
quantitative yield.
##STR00167##
A solution of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid (261.2 mg, 0.653
mmol), 26c (108.4 mg, 0.544 mmol) and 4-dimethylaminopyridine (79.7
mg, 0.653 mmol) in dichloromethane (10 mL) was treated with
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (166.9
mg, 0.870 mmol). After stirring at RT for 17 h, the volatiles were
removed in vacuo and the residue was purified by silica gel
chromatography using a 50 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 1:4 to 0:1
to afford the title compound (139.6 mg, 44%) as a white solid.
##STR00168##
A cooled (0.degree. C.) solution of 26d (139.6 mg, 0.240 mmol) in
anhydrous dichloromethane (10 mL) was treated with trimethylsilyl
methanesulfonate (90 .mu.L, 0.480 mmol). After stirring for 1.5 h
at 0.degree. C., the reaction was quenched with saturated sodium
bicarbonate. The aqueous layer was extracted with dichloromethane.
The organics were combined, filtered through a phase separator and
the volatiles were removed in vacuo to provide the intermediate
amine. A solution of this amine, (E)-(R)-2-methyl-pent-3-enoic acid
(32.9 mg, 0.288 mmol) and 1-hydroxybenzotriazole (38.9 mg, 0.288
mmol) in acetonitrile (10 mL) was treated with
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (64.4
mg, 0.336 mmol). After stirring at RT for 17.5 h the volatiles were
removed in vacuo and the residue was purified by silica gel
chromatography using a 25 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 0:1 to
afford the title compound (69.8 mg, 50%) as a colorless oil.
##STR00169##
Compound 26 was prepared in the same manner as compound 25 using
26e instead of
(S)-1-{(S)-2-[(S)-2-((2R,3R)-3-methoxy-2-methyl-hept-6-enoylamino)-3-meth-
yl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylic acid
methyl-((R)-6-vinyl-indan-1-yl)-amide in 4% yield. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta.0.94-1.00 (m, 6H), 1.47 (d, J=7.6 Hz, 3H),
1.63 (d, J=7.1 Hz, 3H), 1.67-1.80 (m, 5H), 1.85-2.07 (m, 3H), 2.76
(td, J=12.1, 3.3 Hz, 1H), 3.79-3.86 (m, 1H), 4.28 (d, J=10.5 Hz,
1H), 4.36-4.46 (m, 1H), 5.73 (q, J=7.1 Hz, 1H), 5.93 (q, J=6.9 Hz,
1H), 6.32-6.51 (m, 2H), 7.41 (d, J=8.5 Hz, 1H), 7.65 (s, 1H),
7.77-7.85 (m, 2H), 8.21 (d, J=8.5 Hz, 1H). LCMS (m/z) 536.1 [M+H],
Tr=1.80 min.
Example 27
##STR00170##
A cooled (-78.degree. C.) solution of (of
(R)-4-benzyl-3-propionyl-oxazolidin-2-one (3.00 g, 12.9 mmol) in
anhydrous tetrahydrofuran (40 mL) was treated with potassium
bis(trimethylsilyl)amide (19.3 mL, 19.3 mmol, 1 M in
tetrahydrofuran). After stirring for 0.45 h at -78.degree. C., the
mixture was treated with allyl bromide (5.6 mL, 64.3 mmol). After
stirring for 2 h at -40.degree. C., the reaction was quenched with
2 M hydrochloric acid. The aqueous was extracted with ethyl acetate
(2.times.50 mL). The organics were combined, and the volatiles were
removed in vacuo. The residue was purified by silica gel
chromatography using a 50 g Biotage cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 1:1 to
afford the title compound (2.73 g, 78%) as a colorless oil.
##STR00171##
A solution of 27a (2.73 g, 9.97 mmol) in ethanol/water (22 mL,
10:1) was treated with rhodium(III) chloride hydrate (52 mg, 0.25
mmol). After stirring at 85.degree. C. for 3 h, the reaction
mixture was cooled to RT and the volatiles were removed in vacuo.
The residue was purified by silica gel chromatography using a 50 g
Biotage cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 4:1 to afford the title compound
(1.76 g, 65%) as a colorless oil.
##STR00172##
A cooled (0.degree. C.) solution of 27b (1.76 g, 6.44 mmol) in
tetrahydrofuran/water (60 mL, 2:1) was subsequently treated with
hydrogen peroxide (3.3 mL, 32.2 mmol, 30% in water) and lithium
hydroxide hydrate (534 mg, 12.9 mmol). After stirring for 2 h at
0.degree. C., the reaction was quenched with sodium metabisulfite
(12.2 g, 64.4 mmol). After stirring at RT for 1 h, the mixture was
acidified with 2 M hydrochloric acid and the aqueous layer was
extracted with dichloromethane (2.times.100 mL). The organics were
combined, filtered and the volatiles were removed in vacuo. The
residue was purified by silica gel chromatography using a 50 g
Biotage cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 4:1 to afford the title compound
(463 mg, 63%) as a colorless oil.
##STR00173##
A cooled (0.degree. C.) solution of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid
(R)-1-(7-vinyl-quinolin-2-yl)-ethyl ester (380 mg, 0.65 mmol) in
anhydrous dichloromethane (10 mL) was treated with trimethylsilyl
methanesulfonate (237 .mu.L, 1.31 mmol). After stirring for 1 h at
0.degree. C., the reaction was quenched with saturated sodium
bicarbonate. The aqueous layer was extracted with dichloromethane.
The organics were separated and combined, volatiles were removed in
vacuo to provide the intermediate amine (305 mg). A solution of
this amine, 27c (90 mg, 0.76 mmol) and 1-hydroxybenzotriazole (103
mg, 0.76 mmol) in acetonitrile (20 mL) was treated with
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (170
mg, 0.89 mmol). After stirring at RT for 16 h the volatiles were
removed in vacuo and the residue was purified by silica gel
chromatography using a 25 g Biotage cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 0:1 to
afford the title compound (236 mg, 65%) as a colorless oil.
##STR00174##
Compound 27 was prepared in the same manner as compound 25 using
27d instead of
(S)-1-{(S)-2-[(S)-2-((2R,3R)-3-methoxy-2-methyl-hept-6-enoylamino)-3-meth-
yl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylic acid
methyl-((R)-6-vinyl-indan-1-yl)-amide in 5% yield. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta.0.85-1.00 (m, 6H), 1.30 (d, J=6.9 Hz, 3H),
1.58 (d, J=7.1 Hz, 3H), 1.62-1.80 (m, 6H), 1.83-2.07 (m, 3H),
2.70-2.82 (m, 1H), 3.38-3.48 (m, 1H), 3.76-3.84 (m, 1H), 4.20-4.28
(m, 1H), 4.36-4.47 (m, 1H), 5.62-5.74 (m, 1H). 5.93 (q, J=6.9 Hz,
1H), 6.30-6.51 (m, 2H), 7.39 (d, J=8.5 Hz, 1H), 7.63 (s, 1H),
7.70-7.80 (m, 2H), 8.13-8.22 (m, 1H). LCMS (m/z) 536.2 [M+H],
Tr=2.15 min.
Example 28
##STR00175##
Potassium carbonate (647 mg, 4.68 mmol) and di-tert-butyl
dicarbonate (716 mg, 3.28 mmol) were added to
2-(3-bromo-phenyl)-piperidine (750 mg, 3.12 mmol) in
dichloromethane (10 mL). After overnight stirring at RT water (20
mL) was added to the solution. The resulting biphasic solution was
separated into organic and aqueous phases. The aqueous phase was
back extracted with dichloromethane (20 mL). The combined organic
phases were dried over anhydrous sodium sulfate, filtered and
concentrated to afford the title compound (1.03 g, 97%) as a pale
yellow oil.
##STR00176##
To a solution of 28a (1.03 g, 3.03 mmol) and potassium
vinyltrifluoroborate (488 mg, 3.64 mmol) in n-propanol (30 mL),
under an atmosphere of nitrogen was added
1,1'bis(diphenylphosphino)ferrocenedichloropalladium(II),
dichloromethane adduct (49 mg, 0.06 mmol) and triethylamine (306
mg, 422 .mu.L, 3.03 mmol). The reaction was heated to reflux and
left to stir for 3 h before cooling to RT. The reaction mixture
poured onto water and the resultant solution was extracted with
diethyl ether (3.times.30 mL). The combined organics were dried
over anhydrous sodium sulfate, filtered and concentrated. The
resultant residue was purified by silica gel chromatography using a
stepwise gradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to afford
the title compound (652 mg, 75%) as a pale yellow oil.
##STR00177##
A solution of
(2R,3R)-1-((1R,5S)-10,10-dimethyl-3,3-dioxo-3lambda*6*-thia-4-aza-tricycl-
o[5.2.1.0*1,5*]dec-4-yl)-3-hydroxy-2-methyl-hept-6-en-1-one (250
mg, 0.703 mmol) in anhydrous dichloromethane (7 mL) was prepared
and trimethyloxonium tetrafluoroborate (208 mg, 1.406 mmol) was
added. The reaction mixture was stirred at RT for 15 h. The
reaction mixture was treated with methanol (1 mL), then 2 M
hydrochloric acid (20 mL) and saturated brine (20 mL). The mixture
was extracted with ethyl acetate (3.times.15 mL) and the extract
was dried over sodium sulfate, filtered and evaporated to give a
yellow gum. The gum was purified by silica gel chromatography using
iso-hexanes/ethyl acetate 4:1 to give the title compound (223 mg,
86%) as a colorless gum.
##STR00178##
A solution of 2 M lithium hydroxide in water (5 mL, 10 mmol) was
added to a stirred solution of 28c (223 mg, 0.60 mmol) in
tetrahydrofuran (15 mL). The stirred mixture was heated to
60.degree. C. for 15 h. The reaction mixture was partially
evaporated before adding 2 M hydrochloric acid (20 mL). The
solution was extracted with ethyl acetate (3.times.15 mL). The
extract was dried over sodium sulfate, filtered and evaporated to
give a yellow gum (209 mg). The gum was purified by silica gel
chromatography using iso-hexanes/ethyl acetate 3:1 to yield the
title compound (68 mg, 66%) as a yellow gum.
##STR00179##
A solution of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methylbutyrylamino)-propio-
nyl]-hexahydropyridazine-3-carboxylic acid 2,2,2-trichloroethyl
ester (316 mg, 0.59 mmol) in anhydrous dichloromethane (10 mL) was
cooled to 0.degree. C. under a nitrogen atmosphere before adding
trimethylsilyl trifluoromethanesulfonate (160 .mu.L, 0.885 mmol).
The reaction mixture was stirred at 0.degree. C. for 2 h before
adding N,N-diisopropylethylamine (413 .mu.l, 2.36 mmol) to afford
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloroethyl ester. The mixture
was evaporated and the residue dissolved with
(2R,3R)-3-methoxy-2-methyl-hept-6-enoic acid (162 mg, 0.94 mmol.)
in acetonitrile (13 mL) and
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (250
mg, 1.32 mmol) and 1-hydroxybenzotriazole (220 mg, 1.32 mmol) were
added. The reaction was stirred at RT for 15 h then evaporated to
give a yellow oil. The oil was purified by silica gel
chromatography using ethyl acetate to give the title compound (425
mg, 77%) as a white solid.
##STR00180##
A solution of 28e (425 mg, 0.725 mmol) in tetrahydrofuran (20 mL)
was prepared and zinc powder (0.48 g, 7.25 mmol) was added followed
by an aqueous solution of ammonium acetate (1 M, 5 mL, 5 mmol). The
reaction mixture was stirred at RT for 15 h. The reaction was
filtered through hyflo-supercel washing through with ethyl acetate.
The mixture was treated with hydrochloric acid (2 M, 30 mL) and the
layers were separated. The aqueous layer was extracted with ethyl
acetate (25 mL). The organic layers were combined, washed with
brine, filtered and evaporated to give a colorless gum (299 mg)
which was used directly without further purification.
##STR00181##
A solution of 2-(3-vinyl-phenyl)-piperidine-1-carboxylic acid
tert-butyl ester (650 mg, 2.26 mmol) in dichloromethane (15 mL) was
cooled to 0.degree. C., before adding trimethylsilyl
trifluoromethanesulfonate (569 .mu.L, 3.39 mmol). The reaction
mixture was stirred at 0.degree. C. for 1 h before adding
N,N-diisopropylethylamine (1.6 mL, 5.24 mmol) to afford the
2-(3-vinyl-phenyl)-piperidine as a yellow solid. The solid was
redissolved, along with 28f (900 mg, 1.98 mmol) in acetonitrile (20
mL). The solution was cooled to 0.degree. C., before adding
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (1.05 g, 2.77 mmol) and
N,N-diisopropylethylamine (1.4 mL, 7.92 mmol). The stirred reaction
mixture was allowed to slowly warm to RT. After 2 h, the solvent
was evaporated and the remaining residue dissolved in ethyl acetate
(30 mL) and washed with water (3.times.30 mL). The organics were
dried over anhydrous sodium sulfate, filtered and concentrated. The
resultant residue was purified by silica gel chromatography using a
stepwise gradient of iso-hexanes/ethyl acetate 1:0 to 0:1 to afford
the title compound (1.25 g, 100%) as a yellow solid.
##STR00182##
To a stirred solution of 28g (1.05 mg, 1.68 mmol) in
1,2-dichloroethane (550 mL) was added Hoveyda-Grubbs 2.sup.nd
generation catalyst (105 mg, 0.168 mmol). The solution was heated
to 84.degree. C. and was left to stir for 2.5 h. The solvent was
evaporated and the remaining residue was purified by silica gel
chromatography using a stepwise gradient of iso-hexanes/ethyl
acetate/acetone 1:0:0 to 0:9:1. This material was then subjected to
a second round of silica gel chromatography using the same gradient
to afford a yellow solid. A final round of silica gel
chromatography using neat ethyl acetate and eluting purely by
gravity afforded the title compound (175 mg, 18%) as a white solid
as a 6:4 mixture of diastereoisomers. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta.0.84 (d, J=6.9 Hz, 3H), 0.91-1.01 (m, 6H),
1.24-1.37 (m, 6H), 1.47-2.03 (m, 14H), 2.06 (s, 2H), 2.14-2.25 (m,
1H), 2.43-2.77 (m, 4H), 3.48 (s, 1H), 3.56 (s, 2H), 3.63-3.86 (m,
1H), 4.00 (app t, J=14.7 Hz, 1H), 4.12-4.17 (m, 1H), 4.53-4.65 (m,
1H), 5.45 (q, J=7.2 Hz, 1H), 6.27-6.50 (m, 2H), 6.99-7.10 (m, 2H),
7.14-7.22 (m, 1H), 7.32-7.38 (m, 1H). LCMS (m/z) 596.4 [M+H],
Tr=2.51 min.
Examples 29 and 30
##STR00183##
Compound 28 (155 mg, 0.26 mmol) was dissolved in a 1:1 mixture of
acetonitrile/water to a concentration of 7.8 mg/mL. This solution
was then eluted through a reverse phase HPLC system fitted with a
Phenomenex Gemini 10.mu. 110 A, 250.times.21.2 mm column using an
isocratic 2:3 acetonitrile/water flow at 20 mL/min. The mixture was
resolved into the 2 separate diastereoisomers. On concentration
each separate diastereoisomer yielded a white solid. The
stereochemistry of each isomer was not determined. First isomer
eluted, Compound 29 (15 mg, 10%). .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta.0.89-1.05 (m, 8H), 1.20-1.44 (m, 10H), 1.51-1.79 (m, 3H),
1.83-1.99 (m, 4H), 2.03-2.27 (m, 3H), 2.33-2.67 (m, 3H), 2.76 (app
t, J=12.3 Hz, 1H), 3.09 (app t, J=12.3 Hz, 1H), 3.22-3.38 (m, 1H),
3.47 (s, 3H), 3.54-3.62 (m, 1H), 4.17 (d, J=11.7 Hz, 1H), 4.59 (d,
J=10.5 Hz, 1H), 5.39-5.53 (m, 1H), 6.01 (s, 1H), 6.09-6.23 (m, 1H),
6.30-6.55 (m, 2H), 6.99-7.23 (m, 4H). LCMS (m/z) 596.4 [M+H],
Tr=2.53 min.
Second isomer eluted, Compound 30 (22 mg, 14%). .sup.1H NMR (300
MHz, CD.sub.3CN) .delta.0.98-1.04 (m, 6H), 1.26-1.38 (m, 7H),
1.62-1.80 (m, 5H), 1.83-2.00 (m, 4H), 2.03-2.14 (m, 1H), 2.16-2.25
(m, 2H), 2.36-2.51 (m, 2H), 2.54-2.66 (m, 1H), 2.68-2.82 (m, 1H),
3.03-3.16 (m, 1H), 3.26-3.34 (m, 1H), 3.48 (s, 3H), 3.64-3.73 (m,
1H), 3.76-3.86 (m, 1H), 4.00 (app t, J=9.0 Hz, 1H), 4.16 (d, J=12.0
Hz, 1H), 4.55-4.66 (m, 1H), 5.41-5.53 (m, 1H), 5.97-6.06 (m, 1H),
6.10-6.22 (m, 1H), 6.35 (s, 1H), 6.38-6.49 (m, 1H), 7.00-7.11 (m,
2H), 7.14-7.23 (m, 3H). LCMS (m/z) 596.3 [M+H], Tr=2.49 min.
Example 31
##STR00184##
A cooled (0.degree. C.) solution of
(R)-4-benzyl-3-[(R)-2-((S)-1-hydroxy-ethyl)-but-3-enoyl]-oxazolidin-2-one
(233.6 mg, 0.807 mmol, prepared as described in Org. Lett. 2007, 9,
1635-1638) and imidazole (241.7 mg, 3.551 mmol) in
N,N-dimethylformamide (2 mL) was treated with
tert-butyldimethylsilyl chloride (158.2 mg, 1.049 mmol). After
stirring for 24 h at RT, the reaction was quenched with saturated
ammonium chloride. The aqueous layer was extracted with diethyl
ether (2.times.20 mL). The organics were combined, dried over
sodium sulfate, filtered and the volatiles were removed in vacuo.
The residue was purified by silica gel chromatography using a 25 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 4:1 to afford the title compound
(294.6 mg, 90%) as a colorless oil.
##STR00185##
A solution of 31a (294.6 mg, 0.730 mmol),
(R)-1-(7-bromo-quinolin-2-yl)-ethanol (184.0 mg, 0.730 mmol),
palladium(II)acetate (32.8 mg, 0.146 mmol), tri-(o-toluyl)phosphine
(44.4 mg, 0.146 mmol) in anhydrous 1,4-dioxane (10 mL) was treated
with N,N-dicylohexylmethylamine (250 .mu.L, 1.168 mmol). After
stirring at 100.degree. C. for 5 h, the reaction was cooled to RT,
diluted with dichloromethane and saturated sodium bicarbonate. The
aqueous layer was extracted with dichloromethane. The organics were
combined, filtered through a phase separator and the volatiles were
removed in vacuo. The residue was purified by silica gel
chromatography using a 25 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 7:3 to
afford the title compound (230.8 mg, 55%) as a colorless oil.
##STR00186##
A cooled (0.degree. C.) solution of 31b (230.8 mg, 0.401 mmol) in
tetrahydrofuran/water (15 mL, 2:1) was subsequently treated with
hydrogen peroxide (30% aqueous, 210 .mu.L, 2.005 mmol) and lithium
hydroxide hydrate (33.7 mg, 0.803 mmol). After stirring for 2 h at
0.degree. C., the reaction was quenched with sodium metabisulfite
(765 mg, 4.1 mmol). After stirring for 3.5 h at RT the volatiles
were removed in vacuo. The mixture was then diluted with water and
the pH was adjusted with potassium carbonate. The aqueous layer was
washed with dichloromethane (2.times.20 mL) and acidified with 2 M
hydrochloric acid (pH .about.1) then extracted with dichloromethane
(3.times.30 mL). All the organics were combined, dried over sodium
sulfate, filtered and the volatiles were removed in vacuo. The
residue was purified by silica gel chromatography using a 25 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 0:1 to afford the title compound
(69.8 mg, 42%) as a white solid.
##STR00187##
Compound 31d was prepared in the same manner as 22e using 31c
instead of
(E)-4-[2-((R)-1-acetoxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enoic
acid in 46% yield.
##STR00188##
Compound 31 was prepared in the same manner as Compound 22 using
31d instead of
(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-hydroxy-ethyl)-quinolin-7-yl]-2,2-di-
methyl-but-3-enoylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyrid-
azine-3-carboxylic acid 2,2,2-trichloro-ethyl ester in 10% yield.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta.1.00 (d, J=6.7 Hz, 3H),
1.01 (d, J=6.7 Hz, 3H), 1.22-1.44 (m, 4H), 1.64 (d, J=7.1 Hz, 3H),
1.68-1.77 (m, 4H), 1.89-2.08 (m, 3H), 2.71-2.84 (m, 1H), 3.78-3.86
(m, 1H), 4.11 (dd, J=8.2, 6.2 Hz, 1H), 4.17-4.25 (m, 2H), 4.38-4.47
(m, 1H), 5.68 (q, J=7.1 Hz, 1H), 5.94 (q, J=6.9 Hz, 1H), 6.47 (d,
J=16.5 Hz, 1H), 6.69 (dd, J=16.5, 5.3 Hz, 1H), 7.40 (d, J=8.5 Hz,
1H), 7.68 (s, 1H), 7.76 (dd, J=8.7, 1.3 Hz, 1H), 7.81 (d, J=8.7 Hz,
1H), 8.21 (d, J=8.5 Hz, 1H). LCMS (m/z) 566.1 [M+H], Tr=1.65
min.
Example 32
##STR00189##
To a cooled (0.degree. C.) solution of ethyl propiolate (2 mL,
19.735 mmol) in dichloromethane (50 mL) was added dropwise
morpholine (1.7 mL, 19.435 mmol). After stirring at RT for 1.5 h,
the volatiles were removed in vacuo and the residue was purified by
silica gel chromatography using a 50 g Isolute cartridge eluted
with dichloromethane/methanol 20:1 to afford the title compound
(3.5034 g, 97%) as a colorless oil.
##STR00190##
A cooled (0.degree. C.) solution of 4-bromoaniline (2.7 g, 15.7
mmol) in water (30 mL) was subsequently treated with concentrated
hydrochloric acid (3.5 mL) and sodium nitrite (1.3 g, 18.840 mmol).
After 20 min at 0.degree. C., concentrated hydrochloric acid (5.3
mL) and sodium tetrafluoroborate (6.9 g, 62.847 mmol) were added.
After 40 min at 0.degree. C., the intermediate diazonium was
filtered, washed with water, methanol and diethyl ether (2.1021 g)
and was used without further purification. A solution of the
diazonium (7.762 mmol) in acetonitrile (50 mL) was treated with 32a
(1.6614 g, 8.970 mmol). After 1 h at RT, silica gel was added.
After stirring at RT for 16 h, the volatiles were removed in vacuo
and the residue was purified by silica gel chromatography using a
100 g Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 9:1 to afford the title compound
(1.9811 g, 85%) as a highly coloured solid and as a mixture of
tautomers.
##STR00191##
A solution of 32b (1.9811 g, 6.623 mmol) in concentrated sulphuric
acid (25 mL) was heated at 100.degree. C. for 3 h. After cooling to
0.degree. C., the mixture was diluted with water (150 mL) and a
brown solid was filtered off. The filtrate was extracted with
diethyl ether (2.times.50 mL), dichloromethane (2.times.50 mL) and
ethyl acetate (50 mL). The organics were combined, dried over
sodium sulfate, filtered and the volatiles were removed in vacuo to
provide the title compound (1.5094 g, 90%) as an orange solid that
turned dark upon standing.
##STR00192##
A cooled (0.degree. C.) solution of 32c (1.5094 g, 5.964 mmol),
N,O-dimethylhydroxylamine hydrochloride (755.9 mg, 7.753 mmol) and
N,N-diisopropylethylamine (4.2 mL, 23.856 mmol) in acetonitrile (50
mL) was treated with
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (3.175 g, 8.350 mmol). After
stirring for 6 h at RT, the mixture was cooled to 0.degree. C. and
quenched with 1 M hydrochloric acid (60 mL). The aqueous layer was
extracted with ethyl acetate (3.times.50 mL). The organics were
combined, dried over sodium sulfate, filtered and the volatiles
were removed in vacuo. The residue was purified by silica gel
chromatography using a 100 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 1:1 to
afford the title compound (1.4229 g, 81%) as a bright yellow
solid.
##STR00193##
A cooled (-78.degree. C.) solution of 32d (1.4229 g, 4.805 mmol) in
tetrahydrofuran (50 mL) was treated with methylmagnesium chloride
(3.2 mL, 9.610 mmol, 3 M in diethyl ether). After 1 h at
-78.degree. C. and 3 h at 0.degree. C., the reaction was quenched
with saturated ammonium chloride (30 mL). The aqueous layer was
extracted with ethyl acetate (3.times.50 mL). The organics were
combined, dried over sodium sulfate, filtered and the volatiles
were removed in vacuo. The residue was purified by silica gel
chromatography using a 100 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to
afford the title compound (751.5 mg, 62%) as a yellow solid.
##STR00194##
A solution of 32e (751.5 mg, 2.993 mmol) in tetrahydrofuran (30 mL)
was treated with (S)-(-)-2-methyl-CBS-oxazaborolidine (3.6 mL,
3.592 mmol, 1 M in toluene). After 10 min at RT the mixture was
cooled to -60.degree. C. and treated with borane-tetrahydrofuran
complex (6 mL, 5.986 mmol, 1 M in tetrahydrofuran). After 1.5 h at
-55.degree. C. to -30.degree. C., the reaction was quenched with
methanol (20 mL). After stirring at RT for 16 h, the volatiles were
removed in vacuo. The residue was purified by silica gel
chromatography using a 50 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 3:2 to
afford the title compound (345.0 mg, 45%) as a yellow solid.
##STR00195##
A solution of 32f (345.0 mg, 1.363 mmol), palladium II acetate
(61.2 mg, 0.273 mmol), tri-(o-toluyl)phosphine (83.1 mg, 0.273
mmol), 3-butenoic acid tert-butyl ester (560 .mu.L, 3.407 mmol) and
N,N-dicyclohexylmethylamine (470 .mu.L, 2.181 mmol) in anhydrous
1,4-dioxane (20 mL) was heated at 100.degree. C. for 1.7 h. After
cooling to RT the volatiles were removed in vacuo. The residue was
purified by silica gel chromatography using a 50 g Isolute
cartridge eluted with a continuous gradient of iso-hexanes/ethyl
acetate 1:0 to 2:3 to afford the title compound (244.1 mg, 57%) as
a yellow solid.
##STR00196##
A cooled (0.degree. C.) solution of 1e (422.0 mg, 0.793 mmol) in
tetrahydrofuran/water (25 mL, 4:1) was treated with lithium
hydroxide hydrate (67.0 mg, 1.587 mmol). After stirring at
0.degree. C. for 1.5 h, the reaction was quenched with 1 M
hydrochloric acid (20 mL). The aqueous layer was extracted with
ethyl acetate (2.times.30 mL). The organics were combined, dried
over sodium sulfate, filtered and the volatiles were removed in
vacuo then residual trichloroethanol was azeotroped off with
toluene (3.times.) to give the intermediate acid as a white solid
which was then combined with 32g (244.1 mg, 0.793 mmol),
4-dimethylaminopyridine (97.0 mg, 0.793 mmol),
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (243.3
mg, 1.269 mmol) and dichloromethane (20 mL). After stirring at RT
for 16 h, the reaction was quenched with dilute hydrochloric acid.
The aqueous layer was extracted with dichloromethane (30 mL). The
organics were combined and filtered through a phase separator and
the volatiles were removed in vacuo. The residue was purified by
silica gel chromatography using a 50 g Isolute cartridge eluted
with a continuous gradient of iso-hexane/sethyl acetate 1:0 to 1:2
to afford the title compound (164.4 mg, 30% over 2 steps) as a
yellow glass.
##STR00197##
A cooled (0.degree. C.) solution of 32h (164.4 mg, 0.236 mmol) in
anhydrous dichloromethane (20 mL) was treated with trimethylsilyl
methanesulfonate (170 .mu.L, 0.944 mmol). After stirring for 1 h at
0.degree. C., the reaction was quenched with
N,N-diisopropylethylamine (330 .mu.L, 1.888 mmol) and the volatiles
were removed in vacuo. A cooled (0.degree. C.) solution of the
crude amino acid in acetonitrile (80 mL) was subsequently treated
with N,N-diisopropylethylamine (330 .mu.L, 1.888 mmol) and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (125.6 mg, 0.330 mmol). After
stirring at RT for 2.5 h, the volatiles were removed in vacuo. The
residue was purified by silica gel chromatography using a 50 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 0:1 to afford the final compound
(170.7 mg) as a mixture. Reverse phase preparative HPLC which was
eluted with a gradient of water/acetonitrile 95:5 to 0:100 provided
the title compound (30.6 mg, 25%) as a white solid. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta.1.00 (d, J=6.7 Hz, 3H), 1.03 (d, J=6.7
Hz, 3H), 1.61 (d, J=7.3 Hz, 3H), 1.66-2.07 (m, 8H), 2.68-2.81 (m,
1H), 2.97-3.07 (m, 1H), 3.35-3.47 (m, 1H), 3.76-3.89 (m, 1H), 4.28
(d, J=9.4 Hz, 1H), 4.37-4.47 (m, 1H), 4.53-4.67 (m, 1H), 5.52 (q,
J=7.1 Hz, 1H), 6.36-6.51 (m, 2H), 6.72 (d, J=16.0 Hz, 1H), 7.61 (s,
1H), 7.97 (dd, J=8.9, 1.6 Hz, 1H), 8.06 (s, 1H), 8.38 (d, J=8.9 Hz,
1H). LCMS (m/z) 523.1 [M+H], Tr=1.65 min.
Examples 33 and 34
##STR00198##
A cooled (0.degree. C.) solution of
(S)-tetrahydro-pyridazine-1,2,3-tricarboxylic acid
1,2-di-tert-butylester 3-(2,2,2-trichloro-ethyl)ester (5.5297 g,
11.975 mmol) in tetrahydrofuran/methanol (50 mL, 1:1) was treated
with tetrabutylammonium fluoride (23.9 mL, 23.950 mmol, 1 M in
tetrahydrofuran). After stirring at RT for 24 h, the volatiles were
removed in vacuo. The residue was dissolved in diethyl ether and
saturated sodium bicarbonate. The aqueous layer was extracted with
diethyl ether (50 mL). The organics were combined, dried over
sodium sulfate, filtered and the volatiles were removed in vacuo.
The residue was purified by silica gel chromatography using a 100 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 9:1 where the mixed fractions were
further purified by silica gel chromatography using a 50 g Isolute
cartridge eluted with a continuous gradient of iso-hexanes/ethyl
acetate 1:0 to 4:1 to afford the title compound (3.9765 g, 96%) as
a colorless oil and as a mixture of rotamers.
##STR00199##
A cooled (-78.degree. C.) solution of 33a (1.4112 g, 4.097 mmol) in
anhydrous tetrahydrofuran (10 mL) was treated with lithium
bis(trimethylsilyl)amide (6.2 mL, 6.146 mmol, 1 M in
tetrahydrofuran). After stirring for 1.25 h at -78.degree. C., the
mixture was treated with iodomethane (640 .mu.L, 10.242 mmol).
After stirring for 1 h at -78.degree. C., 1 h at 0.degree. C. and 1
h at RT, the reaction was quenched with pH 7 buffer at 0.degree. C.
The aqueous was extracted with dichloromethane (2.times.50 mL). The
organics were combined, dried over sodium sulfate, filtered and the
volatiles were removed in vacuo. The residue was purified by silica
gel chromatography using a 50 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to
afford the title compound (760.7 mq, 52%) as a colorless oil and as
a mixture of rotamers.
##STR00200##
A cooled (0.degree. C.) solution of
(S)-2-tert-butoxycarbonylamino-3-methyl-butyric acid (3.020 g,
13.908 mmol), (S)-2-amino-propionic acid methyl ester hydrochloride
(1.9413 g, 13.908 mmol) and N,N-diisopropylethylamine (17.1 mL,
55.632 mmol) in acetonitrile (50 mL) was treated with
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (7.403 g, 19.471 mmol). After
stirring at RT for 6 h, the reaction was quenched with hydrochloric
acid (1 M, 100 mL). The aqueous layer was extracted with ethyl
acetate (2.times.100 mL). The organics were combined, dried over
sodium sulfate, filtered and the volatiles were removed in vacuo.
The residue was purified by silica gel chromatography using a 100 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 3:2 to afford the title compound
(4.0996 g, 97%) as a white solid.
##STR00201##
A cooled (0.degree. C.) solution of 33c (317.0 mg, 1.048 mmol) in
tetrahydrofuran/water (12 mL, 5:1) was treated with lithium
hydroxide hydrate (88.0 mg, 2.096 mmol). After stirring at
0.degree. C. for 2.5 h, the reaction was quenched with hydrochloric
acid (1 M, 30 mL). The aqueous layer was extracted with
dichloromethane (2.times.30 mL). The organics were combined,
filtered through a phase separator and the volatiles were removed
in vacuo to provide crude acid as a white solid.
##STR00202##
A cooled (0.degree. C.) solution of 33b (376.0 mg, 1.049 mmol) in
anhydrous dichloromethane (15 mL) was treated with trifluoroacetic
acid (5 mL). After stirring at 0.degree. C. for 30 min and RT for 2
h, trifluoroacetic acid (4 mL) was added. After 1 h, the volatiles
were removed in vacuo and the residual trifluoroacetic acid
azeotroped off with toluene (3.times.) to provide the
bis-trifluoroacetic acid ammonium salt as an off-white solid. A
cooled (0.degree. C.) solution of the bis-trifluoroacetic acid
ammonium salt and crude
(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propionic
acid as prepared in the previous step in anhydrous dichloromethane
(15 mL) was subsequently treated with N-methylmorpholine (580
.mu.L, 5.240 mmol) and
O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (596.2 mg, 1.572 mmol). After stirring for 19 h
at RT, the reaction was quenched with hydrochloric acid (1 M, 30
mL). The aqueous was extracted with dichloromethane (2.times.30
mL). The organics were combined, washed with saturated aqueous
sodium bicarbonate and filtered through a phase separator. The
volatiles were removed in vacuo and the residue was purified by
silica gel chromatography using a 50 g Isolute cartridge eluted
with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to 1:2
to afford the title compound (373.1 mg, 83% over 2 steps) as a
white foam and as a mixture of diastereoisomers.
##STR00203##
A cooled (0.degree. C.) solution of 33e (373.1 mg, 0.871 mmol) in
anhydrous dichloromethane (10 mL) was treated with trimethylsilyl
methanesulfonate (310 .mu.L, 1.741 mmol). After stirring for 1 h at
0.degree. C., the reaction was quenched with
N,N-diisopropylethylamine (610 .mu.L, 3.484 mmol) and the volatiles
were removed in vacuo. A cooled (0.degree. C.) solution of the
crude amine and (E)-(2R,3R)-3-methoxy-2-methyl-hex-4-enoic acid
(137.8 mg, 0.871 mmol) in anhydrous acetonitrile (15 mL) was
subsequently treated with N,N-diisopropylethylamine (610 .mu.L,
3.484 mmol) and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl
uronium hexafluorophosphate methanaminium (463.7 mg, 1.219 mmol).
After stirring at RT for 18 h, the reaction was quenched with
hydrochloric acid (1 M, 30 mL). The aqueous layer was extracted
with ethyl acetate (2.times.30 mL). The organics were combined,
washed with saturated sodium bicarbonate (20 mL), dried over sodium
sulfate, filtered and the volatiles were removed in vacuo. The
residue was purified by silica gel chromatography using a 25 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 1:2 to 1:4 to afford the title
compound (332.2 mg, 81%) as a colorless solid and as a mixture of
diastereoisomers.
##STR00204##
Compound 33g was prepared in the same manner as
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid
(R)-1-[6-((E)-3-tert-butoxycarbonyl-propenyl)-cinnolin-3-yl]-ethyl
ester using 33f and (R)-1-(3-vinyl-isoquinolin-6-yl)-ethanol
instead of 1e and
(E)-4-[3-((R)-1-hydroxy-ethyl)-cinnolin-6-yl]-but-3-enoic acid
tert-butyl ester in 30% yield over 2 steps.
##STR00205##
A solution of 33g (136.2 mg, 0.214 mmol) in toluene (70 mL) was
degassed by bubbling N.sub.2 gas through for 20 min. Hoveyda-Grubbs
2.sup.nd generation catalyst (27 mg, 0.043 mmol) was added and the
mixture was refluxed for 2.5 h. More Hoveyda-Grubbs 2.sup.nd
generation catalyst (20 mg) was added and after stirring at reflux
for 1 h, the mixture was cooled to RT, the volatiles were removed
in vacuo and the residue was purified by silica gel chromatography
using a 25 g Isolute cartridge eluted with a continuous gradient of
ethyl acetate/acetone 1:0 to 10:1 to afford both diastereoisomers
in separate mixtures. Both mixtures were purified by silica gel
chromatography using a 10 g Isolute cartridge eluted by gravity
with a continuous gradient of ethyl acetate/acetone 1:0 to 10:1 to
provide the more polar diastereoisomer (19.7 mg, 15%) as a white
solid. The less polar diastereomer was further purified by
preparative TLC eluted with ethyl acetate (4 elutions) and
subsequent preparative TLC eluted with iso-hexanes/acetone 7:3 (2
elutions) to afford the title compound (7.6 mg, 6%) as a white
solid. Relative stereochemistry was not assigned.
More polar diastereoisomer Compound 33: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta.0.83-1.07 (m, 7H), 1.41 (d, J=7.1 Hz, 3H), 1.47
(d, J=6.9 Hz, 3H), 1.50 (s, 3H), 1.70 (d, J=6.7 Hz, 3H), 1.75-1.90
(m, 2H), 1.96-2.15 (m, 3H), 2.64 (dd, J=7.6, 3.1 Hz, 1H), 2.70-2.84
(m, 1H), 3.42 (s, 3H), 3.90 (dd, J=8.7, 3.1 Hz, 1H), 4.00 (app t,
J=8.0 Hz, 1H), 4.10 (s, 1H), 4.58-4.69 (m, 1H), 6.00 (dd, J=9.4,
7.1 Hz, 1H), 6.22 (q, J=6.2 Hz, 1H), 6.51 (d, J=9.8 Hz, 1H), 6.93
(s, 1H), 7.13-7.23 (m, 1H), 7.34-7.43 (m, 1H), 7.90-7.97 (m, 2H),
8.47 (s, 1H). LCMS (m/z) 594.3 [M+H], Tr=1.87 min.
Less polar diastereoisomer Compound 34: .sup.1H NMR (300 MHz,
CD.sub.3CN) .delta.-0.51--0.39 (m, 5H), -0.30 (d, J=6.9 Hz, 3H),
0.06 (d, J=7.1 Hz, 3H), 0.15 (d, J=6.9 Hz, 3H), 0.19 (s, 3H),
0.25-0.34 (m, 4H), 0.38-0.45 (m, 1H), 0.81-1.02 (m, 2H), 1.14-1.42
(m, 2H), 2.69 (dd, J=8.7, 2.9 Hz, 1H), 3.00-3.10 (m, 2H), 3.45 (s,
3H), 4.32 (app pentet, J=7.1 Hz, 1H), 4.76 (q, J=6.7 Hz, 1H), 5.53
(d, J=16.3 Hz, 1H), 5.85 (dd, J=16.1, 8.9 Hz, 1H), 6.20 (dd, J=8.5,
1.6 Hz, 1H), 6.41 (d, J=8.9 Hz, 1H), 6.65-6.75 (m, 2H), 6.91 (s,
1H), 7.11 (d, J=7.3 Hz, 1H), 7.79 (s, 1H). LCMS (m/z) 594.3 [M+H],
Tr=1.83 min.
Example 35
##STR00206##
A cooled (0.degree. C.) solution of 2-fluoro-5-hydroxy-benzoic acid
(1.0051 g, 6.438 mmol), N,O-dimethylhydroxylamine hydrochloride
(1.2560 g, 12.856 mmol) and triethylamine (3.6 mL, 25.752 mmol) in
dichloromethane (35 mL) was treated with
O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (3.663 g, 9.658 mmol). After stirring at RT for
20 h, the reaction was quenched at 0.degree. C. with hydrochloric
acid (2 M, 30 mL). The emulsion was filtered on Celite then the
aqueous layer was extracted with dichloromethane (50 mL). The
organics were combined, filtered through a phase separator and the
volatiles were removed in vacuo. The residue was purified by silica
gel chromatography using a 50 g Isolute cartridge eluted with a
continuous gradient of toluene/diethyl ether 1:0 to 1:2 to afford
the title compound (260.5 mg, 20%) as a colorless oil.
##STR00207##
A solution of 35a (260.5 mg, 1.308 mmol) in acetone (20 mL) was
subsequently treated with potassium carbonate (903.8 mg, 6.539
mmol) and allylbromide (340 .mu.L, 3.924 mmol). After stirring at
RT for 24 h, the reaction was quenched with water (20 mL). The
aqueous layer was extracted with dichloromethane (2.times.30 mL).
The organics were combined, filtered through a phase separator and
the volatiles were removed in vacuo. The residue was purified by
silica gel chromatography using a 25 g Isolute cartridge eluted
with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to 3:2
to afford the title compound (289.7 mg, 92%) as a colorless
oil.
##STR00208##
A cooled (-78.degree. C.) solution of 35b (289.7 mg, 1.210 mmol) in
tetrahydrofuran (10 mL) was treated with methylmagnesium chloride
(810 .mu.L, 2.422 mmol, 3 M in diethyl ether). After 1.25 h at
-78.degree. C., 2 h at 0.degree. C. and 16 h at RT, methylmagnesium
chloride (810 .mu.L, 2.422 mmol, 3 M in diethyl ether) was added.
After 2.5 h at -78.degree. C. and 1.2 h at RT, the reaction was
quenched with silica gel and the volatiles were removed in vacuo.
The residue was purified by silica gel chromatography using a 25 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 9:1 to afford the title compound
(196.7 mg, 84%) as a colorless oil.
##STR00209##
A solution of 35c (196.7 mg, 1.013 mmol) in tetrahydrofuran (15 mL)
was treated with (S)-(-)-2-methyl-CBS-oxazaborolidine (1.2 mL,
1.215 mmol, 1 M in toluene). After 10 min at RT the mixture was
cooled to -50.degree. C. and treated with borane tetrahydrofuran
complex (2.1 mL, 2.026 mmol, 1 M in tetrahydrofuran). After 1.5 h
at -50.degree. C. to -40.degree. C., the reaction was quenched with
methanol (6 mL). After stirring at RT for 22 h, the volatiles were
removed in vacuo. The residue was purified by silica gel
chromatography using a 25 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to
afford the title compound (179.4 mg, 90%) as a white solid.
##STR00210##
A cooled (0.degree. C.) solution of 1e (1.9973 g, 3.755 mmol) in
anhydrous dichloromethane (40 mL) was treated with trimethylsilyl
trifluoromethanesulfonate (1.4 mL, 7.510 mmol). After 30 min at
0.degree. C., the reaction mixture was treated with N,
N-diisopropylethylamine (2.6 mL, 15.020 mmol) and the volatiles
were removed in vacuo to afford the corresponding amine. A cooled
(0.degree. C.) solution of this amine,
(E)-(2R,3R)-2-methylhex-6-enoic acid (428.6 mg, 3.755 mmol,
prepared as described in Synlett 2002, 12, pp 2039-2040) and N,
N-diisopropylethylamine (2.6 mL, 15.020 mmol) in acetonitrile (50
mL) was treated with
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (1.999 g, 5.257 mmol). After
stirring at RT for 20 h, the reaction was quenched with
hydrochloric acid (1 M, 100 mL). The aqueous layer was extracted
with ethyl acetate (2.times.50 mL). The organics were combined,
washed with saturated sodium bicarbonate, dried over sodium
sulfate, filtered and the volatiles were removed in vacuo. The
residue was purified by silica gel chromatography using a 50 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/ethyl acetate 1:0 to 1:4 to afford the title compound
(1.6735 g, 84%) as a brown foam.
##STR00211##
A solution of 35e (534.7 mg, 1.013 mmol) in tetrahydrofuran (20 mL)
was subsequently treated with zinc powder (1.457 g, 22.286 mmol)
and a solution of ammonium acetate (1.171 g, 15.195 mmol) in water
(5 mL). After stirring at RT for 24 h, the mixture was filtered
through Celite. The solid was rinsed with saturated potassium
bisulfate and ethyl acetate. The pH of the filtrate was adjusted
with 2 M hydrochloric acid then the aqueous layer was extracted
with ethyl acetate (2.times.50 mL). The organics were combined,
dried over sodium sulfate, filtered and the volatiles were removed
in vacuo. Residual acetic acid was azeotroped off with toluene
(3.times.) to provide the corresponding acid as a white solid. A
solution of the crude acid,
(R)-1-(5-allyloxy-2-fluoro-phenyl)-ethanol (179.4 mg, 0.914 mmol)
and 4-dimethylaminopyridine (123.7 mg, 1.013 mmol) in
dichloromethane (15 mL) was treated with
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (310.8
mg, 1.621 mmol). After stirring at RT for 20 h, the reaction was
quenched at 0.degree. C. with hydrochloric acid (2 M, 15 mL). The
aqueous layer was extracted with ethyl acetate (2.times.). The
organics were combined, dried over sodium sulfate, filtered and the
volatiles were removed in vacuo. The residue was purified by silica
gel chromatography using a 50 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 1:3 to
afford the title compound (310.5 mg, 59% over 2 steps) as a white
solid.
##STR00212##
A solution of 35f (310.5 mg, 0.540 mmol) in dichloroethane (180 mL)
was treated with Hoveyda-Grubbs 2.sup.nd generation catalyst (67.7
mg, 0.108 mmol). After stirring at reflux for 2 h, the reaction was
cooled to RT and the volatiles were removed in vacuo. The residue
was purified by silica gel chromatography using a 50 g Isolute
cartridge eluted with a continuous gradient of iso-hexanes/ethyl
acetate 1:0 to 0:1 then using a 25 g Isolute cartridge eluted by
gravity with a continuous gradient of iso-hexanes/ethyl acetate 1:0
to 0:1 to afford the title compound (109.9 mg) in a mixture.
Purification by preparative TLC eluted with iso-hexanes/acetone 3:1
(3 elutions) followed by silica gel chromatography using a 25 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/acetone 1:0 to 3:2 provided the title compound (58.7
mg, 20%) as a white solid. .sup.1H NMR (300 MHz, CD.sub.3CN)
.delta.0.85-0.97 (m, 6H), 1.17-1.22 (m, 3H), 1.28-1.39 (m, 4H),
1.44-1.63 (m, 6H), 1.70-1.82 (m, 1H), 1.83-1.92 (m, 1H), 2.30-2.45
(m, 3H), 3.62-3.73 (m, 2H), 3.89 (app t, J=8.5 Hz, 1H), 4.28 (d,
J=8.0 Hz, 1H), 4.40-4.60 (m, 2H), 5.32 (app pentet, J=6.9 Hz, 1H),
5.62-5.74 (m, 1H), 5.77-5.89 (m, 1H), 6.02 (q, J=6.7 Hz, 1H), 6.40
(d, J=7.8 Hz, 1H), 6.76-6.85 (m, 2H), 7.01 (app t, J=9.4 Hz, 1H),
7.19 (br s, 1H). LCMS (m/z) 547.2 [M+H], Tr=2.39 min.
Example 36
##STR00213##
To a solution of Compound 35 (44.0 mg, 0.080 mmol) in ethyl acetate
(5 mL) was added palladium on carbon (10%, 5 mg). The atmosphere
was purged of oxygen. After stirring at RT under an atmosphere of
hydrogen for 2.5 h, the volatiles were removed in vacuo and the
residue was purified by silica gel chromatography using a 20 g
Isolute cartridge eluted with a continuous gradient of
iso-hexanes/acetone 1:0 to 3:2 followed by preparative TLC eluted
with iso-hexanes/acetone 3:2 to afford the final compound (19.0 mg,
43%) as a white solid. .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta.0.85-0.99 (m, 8H), 1.14-1.22 (m, 4H), 1.26-1.42 (m, 4H),
1.48 (d, J=6.7 Hz, 1H), 1.52-1.84 (m, 8H), 2.28-2.42 (m, 1H),
3.61-3.72 (m, 1H), 3.75-4.10 (m, 4H), 4.25 (d, J=9.4 Hz, 1H), 5.26
(app pentet, J=7.1 Hz, 1H), 6.02 (q, J=6.7 Hz, 1H), 6.34 (d, J=8.5
Hz, 1H), 6.75-6.86 (m, 2H), 6.95-7.12 (m, 2H). LCMS (m/z) 549.3
[M+H], Tr=2.54 min.
Example 37:
(E)-(2R,5S,11S,14S,17R,18R)-2,14-Diisopropyl-18-methoxy-11,17-di
methyl-22-oxa-3,9,12,15,28-pentaaza-tricyclo[21.3.1.1*5,9]octacosa-1(27),-
19,23,25-tetraene-4,10,13,16-tetraone
##STR00214##
37a was prepared in the same manner as 35b using
3-hydroxybenzaldehyde instead of
2-fluoro-5-hydroxy-N-methoxy-N-methyl-benzamide in 78% yield.
##STR00215##
A cooled (0.degree. C.) suspension of 37a (1.36 g, 8.434 mmol) and
magnesium sulfate (5 g) in dichloromethane (40 mL) was treated with
D-valinol (870.0 mg, 8.434 mmol). After stirring at 0.degree. C. to
RT for 23 h, the mixture was filtered and the volatiles were
removed in vacuo. The residue was dissolved in anhydrous
dichloromethane (40 mL) and was subsequently treated with
triethylamine (1.3 mL, 9.277 mmol) and a solution of trimethylsilyl
chloride (9.3 mL, 9.277 mmol, 1 M in dichloromethane). After
stirring at RT for 24 h, the volatiles were removed in vacuo and
the residue was triturated with diethyl ether/iso-hexane (100 mL,
1:1). The white solid was filtered off and the filtrate was
evaporated to dryness to provide the intermediate imine. In a
cooled (-40.degree. C.) 3-neck round-bottom flask, equipped with a
nitrogen line and an addition funnel was introduced anhydrous
diethyl ether (25 mL) and a solution of iso-propyllithium (29 mL,
20.242 mmol, 0.7 M in pentane). To this mixture, a solution of the
imine in anhydrous diethyl ether (25 mL) was added dropwise over 15
min. After stirring at -40.degree. C. for 2.5 h, the reaction was
quenched with hydrochloric acid (2 M, 50 mL) and the mixture was
allowed to warm to RT. The acidic aqueous layer was basified with
NaOH pellets at 0.degree. C. and then extracted with diethyl ether
(2.times.). The organics were combined, dried over sodium sulfate,
filtered and the volatiles were removed in vacuo. The residue was
purified by silica gel chromatography using a 50 g Isolute
cartridge eluted with a continuous gradient of iso-hexanes/ethyl
acetate 1:0 to 1:1 to afford the title compound (470.1 mg, 19% over
3 steps) as a colorless oil.
##STR00216##
A solution of 37b (470.1 mg, 1.613 mmol) in methanol (10 mL) and
aqueous methylamine (3 mL, 40 wt % in water) was treated with
periodic acid (1.213 g, 5.323 mmol). After stirring for 24 h at RT
more aqueous methylamine (4 mL, 40 wt % in water) and periodic acid
(1.213 g, 5.323 mmol) were added. After stirring for 17 h at RT the
mixture was filtered over Celite and the solid rinsed with
methanol. The volatiles were removed in vacuo and the residue was
partitioned between water and diethyl ether. The aqueous layer was
extracted with diethyl ether, the organics were combined, dried
over sodium sulfate, filtered and the volatiles were removed in
vacuo. The residue was purified by silica gel chromatography using
a 25 g Isolute cartridge eluted with ethyl acetate to afford the
title compound (191.8 mg, 58%) as a light yellow oil.
##STR00217##
A cooled (0.degree. C.) solution of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid (324.4 mg, 0.810
mmol), 37c (166.3 mg, 0.810 mmol) and N,N-diisopropylethylamine
(560 .mu.L, 3.240 mmol) in acetonitrile (15 mL) was treated with
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (431.2 mg, 1.134 mmol). After
stirring at RT for 20 h, the reaction was quenched with
hydrochloric acid (2 M, 25 mL) at 0.degree. C. The aqueous layer
was extracted with ethyl acetate (2.times.). The organics were
combined, dried over sodium sulfate, filtered and the volatiles
were removed in vacuo. The residue was purified by silica gel
chromatography using a 50 g Isolute cartridge eluted with a
continuous gradient of iso-hexanes/ethyl acetate 1:0 to 0:1 to
afford the title compound (230.5 mg, 48%) as a solid.
##STR00218##
Compound 37e was prepared in the same manner as
1-{(S)-2-[(S)-2-((E)-(2R,3R)-3-methoxy-2-methyl-hex-4-enoylamino)-3-methy-
l-butyrylamino]-propionyl}-3-methyl-hexahydro-pyridazine-3-carboxylic
acid methyl ester using 37d instead of
1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propionyl-
]-3-methyl-hexahydro-pyridazine-3-carboxylic acid methyl ester in
30% yield over 2 steps.
##STR00219##
Compound 37 was prepared in the same manner as 25 using 37e instead
of
(S)-1-{(S)-2-[(S)-2-((2R,3R)-3-methoxy-2-methyl-hept-6-enoylamino)-3-meth-
yl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylic acid
methyl-((R)-6-vinyl-indan-1-yl)-amide in 43% yield. .sup.1H NMR
(300 MHz, d.sub.6-DMSO) .delta.0.74-0.91 (m, 12H), 1.15 (d, J=7.1
Hz, 3H), 1.28 (d, J=7.3 Hz, 3H), 1.40-1.64 (m, 3H), 1.71-1.88 (m,
3H), 1.98 (app sextet, J=6.7 Hz, 1H), 2.66-2.75 (m, 2H), 3.38-3.53
(m, 1H), 3.89 (dd, J=6.7, 2.9 Hz, 1H), 4.10 (app t, J=8.9 Hz, 1H),
4.14-4.22 (m, 1H), 4.42 (qd, J=10.0, 4.2 Hz, 1H), 4.57 (app t,
J=8.2 Hz, 1H), 4.76 (d, J=11.8 Hz, 1H), 5.23 (app t, J=7.3 Hz, 1H),
5.69-5.90 (m, 2H), 6.77-6.90 (m, 3H), 7.05 (d, J=9.4 Hz, 1H),
7.16-7.24 (m, 1H), 8.01 (d, J=8.5 Hz, 1H), 8.14 (d, J=8.5 Hz, 1H).
LCMS (m/z) 586.3 [M+H], Tr=2.34 min.
Example 38
##STR00220##
To a mixture of (R)-1-(3-bromo-phenyl)-ethanol (201 mg, 1.00 mmol),
4-(2-ethoxy-2-oxoethoxy)benzeneboronic acid (Acros Organics, 224
mg, 1.00 mmol) in 1,2-dimethoxyethane (4 mL) were added potassium
carbonate (276 mg, 2.00 mmol) and water (1 mL). The mixture was
stirred at RT and tetrakis(triphenylphosphine) palladium(0) (58 mg,
0.05 mmol) was added then the reaction mixture was heated at
100.degree. C. in a microwave reactor for 30 min. The reaction
mixture was then diluted with ethyl acetate and water. The layers
were separated and the organics washed with water and brine, dried
over magnesium sulfate, filtered and evaporated. The residue was
purified by silica gel chromatography using a gradient of
iso-hexanes/ethyl acetate 9:1 to 7:3 to afford the title compound
(230 mg, 76%) as an oil.
##STR00221##
A solution of 38a (230 mg, 0.77 mmol) in tetrahydrofuran (4 mL) was
stirred at 5.degree. C. under nitrogen, a solution of lithium
hydroxide monohydrate (92 mg, 1.54 mmol) was added and the reaction
mixture was stirred at 5.degree. C. for 2 h and then at RT
overnight. The solution was acidified with 2 M hydrochloric acid
and extracted with ethyl acetate (2.times.). The organic extracts
were combined, washed with water and brine, dried over magnesium
sulfate and evaporated to afford the title compound (175 mg, 84%)
as a white solid.
##STR00222##
A solution of 1e (372 mg, 0.70 mmol) in dichloromethane (15 mL) was
cooled in an ice-water bath under nitrogen. Trimethylsilyl
trifluoromethanesulfonate (0.18 mL, 1.05 mmol) was added dropwise,
and the resulting solution was stirred for 1 hour. Cold saturated
aqueous sodium hydrogen carbonate solution (15 mL) was added and
the mixture was stirred at 0.degree. C. for 15 min. The organic
layer was separated, washed with brine, dried over magnesium
sulfate and evaporated to afford
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (0.70 mmol)
which was used without further purification. A solution of
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (0.70 mmol)
in acetonitrile (15 mL) was stirred at 0.degree. C. under nitrogen.
38b (175 mg, 0.64 mmol) and 1-hydroxybenzotriazole hydrate (123 mg,
0.64 mmol, wetted with not less than 20 wt. % water) were added,
followed by N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
hydrochloride (173 mg, 0.90 mmol) and the reaction mixture was
stirred at RT for 2 h. The solvent was evaporated. The residue was
dissolved in ethyl acetate and the solution was washed with water
(3.times.) followed by brine, dried over magnesium sulfate and
evaporated. The residue was purified by silica gel chromatography
using a gradient of iso-hexanes/ethyl acetate 1:2 to 0:1. The
product was triturated with diethyl ether and dried to afford the
title compound (367 mg, 83%) as a white solid.
##STR00223##
A solution of 38c (343 mg, 0.50 mmol) in tetrahydrofuran (10 mL)
was stirred at RT under nitrogen. Zinc powder (715 mg, 11 mmol) was
added followed by a solution of ammonium acetate (578 mg, 7.50
mmol) in water (5 mL). The reaction mixture was stirred at RT under
nitrogen for 70 h. The reaction mixture was filtered through Celite
and the filter pad was washed with ethyl acetate and 2 M aqueous
hydrochloric acid. The filtrate was acidified to pH 2 with 2 M
aqueous hydrochloric acid, solid sodium chloride was added to
saturate the aqueous layer and the mixture was extracted with ethyl
acetate. The ethyl acetate extracts were combined, washed with
brine, dried over magnesium sulfate and evaporated. The residue was
co-evaporated with toluene (3.times.) to afford the title compound
(237 mg, 86%) as a white powder.
##STR00224##
A solution of 38d (100 mg, 0.18 mmol) in dichloromethane (180 mL)
was stirred at RT under nitrogen. 4-Dimethylaminopyridine (33 mg,
0.27 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
hydrochloride (104 mg, 0.54 mmol) were added and the reaction
mixture was stirred at RT for 18 h. The solvent was evaporated. The
residue was purified by silica gel chromatography using a gradient
of iso-hexanes/ethyl acetate 1:2 to 0:1 followed by silica gel
chromatography using a gradient of ethyl acetate to ethyl
acetate/acetone 4:1. The residue was co-evaporated with
dichloromethane then triturated with diethyl ether to afford a
solid. The solid was washed with ether and dried to afford the
title compound (8 mg, 9%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) 0.95 (d, J=6.2 Hz, 3H), 0.96 (d, J=6.5 Hz, 3H), 1.39
(d, J=7.1 Hz, 3H), 1.54-1.80 (m, 5H), 1.82-2.10 (m, 3H), 2.52-2.63
(m, 1H), 3.29-3.62 (m, 2H), 4.07 (app t, J=9.6 Hz, 1H), 4.45 (br d,
J=13.6 Hz, 1H), 4.67 (ABq, .DELTA..delta..sub.AB=0.12,
J.sub.AB=16.0 Hz, 2H), 4.88-5.01 (m, 1H), 5.93-6.00 (m, 2H), 6.57
(d, J=10.3 Hz, 1H), 6.94 (d, J=8.5 Hz, 2H), 7.15-7.53 (m, 6H). LCMS
(m/z) 537.2 [M+H], Tr=2.34 min.
Example 39
##STR00225##
A solution of 4-iodo phenol (2.2 g, 10.0 mmol),
(R)-2-hydroxy-propionic acid methyl ester (0.95 mL, 10.0 mmol) and
triphenylphosphine (2.62 g, 10.0 mmol) was prepared in
tetrahydrofuran (40 mL). Diisopropyl azodicarboxylate (2.0 mL. 10
mmol) was added dropwise and the reaction was stirred at -5.degree.
C. for 1 hour and then at RT for 2 h. The tetrahydrofuran was
evaporated and diethyl ether/iso-hexanes (1:10, 50 mL) was added.
The mixture was stirred at RT for 10 min where a precipitate
formed. The filtrate was decanted off and evaporated. The residue
was purified by silica gel chromatography using a gradient of
iso-hexanes/ethyl acetate 20:1 to 10:1 to afford the title compound
(2.02 g, 66%) as an oil.
##STR00226##
(R)-1-(3-bromophenyl)-ethanol (1.0 g, 4.97 mmol),
bis(pinacolato)diboron (1.39 g, 5.47 mmol),
1,1'bis(diphenylphosphino)ferrocenedichloropalladium(II),
dichloromethane adduct (203 mg, 0.249 mmol) and potassium acetate
(976 mg, 9.94 mmol) were dissolved in 1,4-dioxane (10 mL) and the
reaction was heated to reflux and left to stir over 3 days. The
reaction was allowed to cool to RT before being filtered through a
pad of Hyflo. The pad was then washed with ethyl acetate and the
combined organics were then concentrated and purified by silica gel
chromatography using a stepwise gradient of iso-hexanes/ethyl
acetate 1:0 to 4:1 to afford the title compound (936 mg, 76%) as a
pale yellow oil.
##STR00227##
A solution of 39b (496 mg, 2.00 mmol) and 39a (612 mg, 2.00 mmol)
in 1,2-dimethoxyethane (4 mL) was stirred at RT under nitrogen. A
solution of 2 M aqueous sodium carbonate (4 mL) was added followed
by tetrakis(triphenylphosphine) palladium(0) (116 mg, 0.1 mmol) and
the reaction mixture was heated at 80.degree. C. for 1 hour. The
reaction mixture was cooled to RT, water was added and the mixture
was extracted with ethyl acetate. The ethyl acetate extracts were
combined, washed with brine, dried over sodium sulfate and
evaporated. The residue was purified by silica gel chromatography
using a gradient of iso-hexanes/ethyl acetate 17:3 to 1:1 to afford
the title compound (253 mg, 42%) as a gum.
##STR00228##
A solution of 39c (250 mg, 0.83 mmol) in tetrahydrofuran (4 mL) was
stirred at 5.degree. C. under nitrogen. A solution of lithium
hydroxide monohydrate (42 mg, 1.00 mmol) in water (1 mL) was added
and the reaction mixture was stirred at 5.degree. C. for 1 hour.
The reaction mixture was acidified to pH 3 with 2 M aqueous
hydrochloric acid and the mixture extracted with ethyl acetate. The
organic extracts were separated, washed with water and brine, dried
over magnesium sulfate and evaporated to afford the title compound
(224 mg, 94%) as a white foam.
##STR00229##
A solution of 1e (478 mg, 0.9 mmol) in dichloromethane (15 mL) was
cooled in an ice-water bath under nitrogen. Trimethylsilyl
trifluoromethanesulfonate (0.25 mL, 1.35 mmol) was added dropwise,
and the resulting solution was stirred for 1 h. Cold saturated
aqueous sodium hydrogen carbonate solution (15 mL) was added and
the mixture was stirred at 0.degree. C. for 15 min. The organic
layer was separated, washed with brine, dried over magnesium
sulfate and evaporated to afford
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (0.9 mmol)
which was used without further purification. A solution of
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (0.9 mmol) in
acetonitrile (15 mL) was stirred at 0.degree. C. under nitrogen.
39d (224 mg, 0.78 mmol) and 1-hydroxybenzotriazole hydrate (184 mg,
0.96 mmol, wetted with not less than 20 wt. % water) were added,
followed by N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
hydrochloride (225 mg, 1.17 mmol) and the reaction mixture was
stirred at RT for 2 h. The solvent was evaporated. The residue was
dissolved in ethyl acetate and the solution was washed with water
(3.times.) followed by brine, dried over magnesium sulfate and
evaporated. The residue was purified by silica gel chromatography
using a gradient of iso-hexanes/ethyl acetate 1:2 to 0:1 to afford
the title compound (490 mg, 90%) as a gum.
##STR00230##
A solution of 39e (490 mg, 0.70 mmol) in tetrahydrofuran (15 mL)
was stirred at RT under nitrogen. Zinc powder (1.00 g, 15.40 mmol)
was added followed by a solution of ammonium acetate (810 mg, 10.50
mmol) in water (8 mL). The reaction mixture was stirred at RT under
nitrogen for 24 h. The reaction mixture was filtered through Celite
and the filter pad was washed with ethyl acetate and 2 M aqueous
hydrochloric acid. The filtrate was acidified to pH 2-3 with 2 M
aqueous hydrochloric acid. Solid sodium chloride was added to
saturate the aqueous layer and the mixture was extracted with ethyl
acetate. The ethyl acetate extracts were combined and washed with
brine. The organic extracts were passed through a hydrophobic frit
and the filtrate was evaporated. The residue was co-evaporated with
ethyl acetate (3.times.) then toluene (3.times.) to afford the
title compound (394 mg, 98%) as a white solid.
##STR00231##
A solution of 39f (100 mg, 0.18 mmol) in dichloromethane (180 mL)
was stirred at RT under nitrogen. 4-Dimethylaminopyridine (44 mg,
0.36 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
hydrochloride (138 mg, 0.72 mmol) were added and the reaction
mixture was stirred at RT for 4 h. The solvent was evaporated.
Ethyl acetate was added to the residue and the mixture was washed
with aqueous citric acid (pH 4) and brine. The organic layer was
dried over magnesium sulfate and the solvent was evaporated. The
residue was purified by silica gel chromatography using a gradient
of iso-hexanes/ethyl acetate 3:7 to 0:1. The residue was triturated
with diethyl ether/iso-hexanes 1:1 to afford the title compound (10
mg, 10%) as a white solid. .sup.1H NMR (300 MHz, CD.sub.3OD) 0.94
(d, J=6.7 Hz, 3H), 0.95 (d, J=6.9 Hz, 3H), 1.51 (d, J=7.1 Hz, 3H),
1.61 (d, J=6.5 Hz, 3H), 1.67 (d, J=6.9 Hz, 3H), 1.72-2.00 (m, 5H),
2.60-2.69 (m, 1H), 3.54-3.63 (m, 1H), 4.13 (d, J=10.5 Hz, 1H), 4.32
(brd, J=12.9 Hz, 1H), 4.57-4.66 (m, 2H), 4.78 (q, J=7.1 Hz, 1H),
5.92 (q, J=6.5 Hz, 1H), 6.91 (d, J=8.9 Hz, 2H), 7.22 (d, J=7.6 Hz,
1H), 7.34-7.61 (m, 5H). LCMS (m/z) 551.2 [M+H], Tr=2.51 min.
Example 40
##STR00232##
40a was prepared in the same manner as
(S)-2-(4-iodo-phenoxy)-propionic acid methyl ester using
(S)-2-hydroxy-propionic acid ethyl ester instead of
(R)-2-hydroxy-propionic acid methyl ester in 28% yield.
##STR00233##
40b was prepared in the same manner as
(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-propionic acid
methyl ester using 40a instead of (S)-2-(4-iodo-phenoxy)-propionic
acid methyl ester in 54% yield.
##STR00234##
40c was prepared in the same manner as
(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-propionic acid
using 40b instead of
(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-propionic acid
methyl ester in 71% yield.
##STR00235##
40d was prepared in the same manner as
(S)-1-[(S)-2-((S)-2-{(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-pr-
opionylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-car-
boxylic acid 2,2,2-trichloro-ethyl ester using 40c instead of
(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-propionic acid in
71% yield.
##STR00236##
40e was prepared in the same manner as
((S)-1-[(S)-2-((S)-2-{(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-p-
ropionylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-ca-
rboxylic acid using 40d instead of
(S)-1-[(S)-2-((S)-2-{(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-pr-
opionylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-car-
boxylic acid 2,2,2-trichloro-ethyl ester in 83% yield.
##STR00237##
A solution of 40e (102 mg, 0.18 mmol) in dichloromethane (180 mL)
was stirred at RT under nitrogen. 4-Dimethylaminopyridine (44 mg,
0.36 mmol) and (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium
hexafluorophosphate (114 mg, 0.22 mmol) were added and the reaction
mixture was stirred at RT for 4 h. The solvent was evaporated.
Ethyl acetate was added to the residue and the mixture was washed
with aqueous citric acid (pH 4) and brine. The organic layer was
separated and dried over magnesium sulfate and evaporated. The
residue was purified by silica gel chromatography using a gradient
of iso-hexanes/ethyl acetate 3:7 to 0:1. The residue was purified
by preparative reverse phase HPLC to afford the title compound (2.8
mg, 3%) as a white solid. .sup.1H NMR (300 MHz, CD.sub.3OD) 1.03
(d, J=6.9 Hz, 3H), 1.05 (d, J=6.9 Hz, 3H), 1.25 (d, J=7.1 Hz, 3H),
1.56 (d, J=6.5 Hz, 3H), 1.57 (d, J=6.3 Hz, 3H), 1.64-2.10 (m, 5H),
2.75-2.83 (m, 1H), 3.60-3.66 (m, 1H), 4.08 (d, J=10.0 Hz, 1H), 4.32
(br d, J=12.9 Hz, 1H), 4.74 (q, J=6.5 Hz, 1H), 5.22 (q, J=7.3 Hz,
1H), 5.98 (q, J=6.5 Hz, 1H), 6.86 (d, J=8.5 Hz, 2H), 7.20-7.25 (m,
1H), 7.39-7.51 (m, 5H). LCMS (m/z) 551.2 [M+H], Tr=2.27 min.
Example 41
##STR00238##
Dichloro (p-cymene) ruthenium(II) dimer (31 mg, 0.05 mmol) and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (44 mg, 0.12
mmol) was suspended in degassed water (20 mL) and the mixture was
degassed with nitrogen for 10 min. The mixture was stirred at
70.degree. C. under nitrogen for 90 min. The resulting yellow
solution was cooled to RT. A solution of
1-(6-bromo-pyridin-2-yl)-ethanone (2.0 mg, 10 mmol) in degassed
tetrahydrofuran (10 mL) and sodium formate (3.4 g, 50 mmol) was
added and the reaction mixture was degassed with nitrogen for 5
min. The reaction mixture was vigorously stirred at 40.degree. C.
for 30 min. The reaction mixture was cooled to RT and was extracted
with ethyl acetate. The organic extracts were combined, washed with
water and brine, dried over sodium sulfate, filtered and
evaporated. The residue was purified by silica gel chromatography
using a gradient of iso-hexanes/ethyl acetate 3:1 to 0:1 to afford
the title compound (1.78 g, 89%) as a brown oil.
##STR00239##
To a mixture of 41a (101 mg, 0.50 mmol),
4-(2-ethoxy-2-oxoethoxy)benzeneboronic acid (112 mg, 0.50 mmol) in
1,2-dimethoxyethane (4 mL) was added a solution of potassium
carbonate (138 mg, 1.00 mmol) in water (1 mL).
Tetrakis(triphenylphosphine)palladium(0) (29 mg, 0.025 mmol) was
added and the reaction mixture was heated at 100.degree. C. in a
microwave reactor for 30 min. The reaction mixture was then diluted
with ethyl acetate and water. 2 M Hydrochloric acid was added to
adjust the pH of the reaction mixture to pH 3 and the volatiles was
evaporated. Methanol was added to the residue and the mixture was
filtered through a hydrophobic frit. The filtrate was evaporated
and the residue was dried in vacuum to afford the title compound
(143 mg, 93%) as a white solid.
##STR00240##
A solution of 1e (240 mg, 0.45 mmol) in dichloromethane (15 mL) was
cooled in an ice-water bath under nitrogen. Trimethylsilyl
trifluoromethanesulfonate (0.12 mL, 0.68 mmol) was added dropwise,
and the resulting solution was stirred for 1 h. Cold saturated
aqueous sodium hydrogen carbonate solution (15 mL) was added and
the mixture was stirred at 0.degree. C. for 15 min. The organic
layer was separated, washed with brine, dried over magnesium
sulfate and evaporated to afford
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (0.45 mmol)
which was used without further purification. A mixture of
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (0.45 mmol),
41b (143 mg, 0.45 mmol) and N,N-diisopropylethylamine (0.16 mL, 0.9
mmol) in acetonitrile (15 mL) was stirred at RT under nitrogen.
1-Hydroxybenzotriazole hydrate (108 mg, 0.56 mmol, wetted with not
less than 20 wt. % water) and
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (130
mg, 0.675 mmol) was added and the reaction mixture was stirred at
RT for 4 h. N,N-Dimethylformamide (2 mL) was added and the reaction
mixture was stirred at RT for 22 h. Additional
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (87
mg, 0.45 mmol) was added and the reaction mixture was stirred at RT
for 6 h. The solvent was evaporated. The residue was suspended in a
mixture of ethyl acetate and aqueous citric acid solution (pH 3)
and the mixture was extracted with ethyl acetate. The organic
extracts were combined, washed with water followed by brine. The
organic layer was filtered through a hydrophobic frit and the
filtrate was evaporated. The residue was purified by silica gel
chromatography using ethyl acetate to afford the title compound (71
mg, 23%) as a white solid.
##STR00241##
A solution of 41c (69 mg, 0.10 mmol) in tetrahydrofuran (2 mL) was
stirred at 0.degree. C. under nitrogen, a solution of lithium
hydroxide monohydrate (5 mg, 0.12 mmol) in water (0.5 mL) was added
and the reaction mixture was stirred at 0.degree. C. for 1 h. The
solution was acidified to pH 3 with 2 M hydrochloric acid and the
solvent was evaporated. The residue was co-evaporated with toluene
(3.times.) to afford
(S)-1-{(S)-2-[(S)-2-(2-{4-[6-((R)-1-hydroxy-ethyl)-pyridin-2-yl]-p-
henoxy}-acetylamino)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazin-
e-3-carboxylic acid (58 mg, 0.1 mmol) as a white solid which was
used crude in the next step. A solution of
(S)-1-{(S)-2-[(S)-2-(2-{4-[6-((R)-1-hydroxy-ethyl)-pyridin-2-yl]-phenoxy}-
-acetylamino)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-car-
boxylic acid (56 mg, 0.1 mmol) in dichloromethane (100 mL) was
stirred at RT under nitrogen. 4-Dimethylaminopyridine (49 mg, 0.4
mmol) was added and the reaction mixture was for stirred for 5 min.
A solution of (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium
hexafluorophosphate (78 mg, 0.15 mmol) in dichloromethane (20 mL)
was added dropwise over 5 min and the reaction mixture was stirred
at RT for 2 h. The solvent was evaporated and the residue was
purified by silica gel chromatography using ethyl acetate. The
residue was purified by preparative reverse phase HPLC to afford
the title compound (14 mg, 26%) as a white solid. .sup.1H NMR (300
MHz, CD.sub.3OD) 0.94 (d, J=6.7 Hz, 3H), 0.98 (d, J=6.5 Hz, 3H),
1.36 (d, J=7.1 Hz, 3H), 1.68 (d, J=6.5 Hz, 3H), 1.72-2.05 (m, 5H),
2.65-2.74 (m, 1H), 3.53-3.60 (m, 1H), 4.14 (d, J=10.7 Hz, 1H),
4.30-4.38 (m, 1H), 4.66 (ABq, .DELTA..delta..sub.AB=0.13,
J.sub.AB=15.8 Hz, 2H), 5.07 (q, J=7.1 Hz, 1H), 5.87 (q, J=6.5 Hz,
1H), 6.89 (d, J=8.9 Hz, 2H), 7.26 (br d, J=7.1 Hz, 1H), 7.71-7.78
(m, 2H), 8.06 (d, J=8.9 Hz, 2H). LCMS (m/z) 538.2 [M+H], Tr=2.07
min.
Example 42
##STR00242##
42a was prepared in the same manner as
(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-propionic acid
methyl ester using 3-(4-bromo-pyrazol-1-yl)-propionic acid methyl
ester instead of (S)-2-(4-iodo-phenoxy)-propionic acid methyl ester
in 28% yield.
##STR00243##
42b was prepared in the same manner as
(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-propionic acid
using 42a instead of
(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-propionic acid
methyl ester in 96% yield.
##STR00244##
42c was prepared in the same manner as compound
(S)-1-[(S)-2-((S)-2-{(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-pr-
opionylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-car-
boxylic acid 2,2,2-trichloro-ethyl ester using 42b instead of
(S)-2-[3'-((R)-1-hydroxy-ethyl)-biphenyl-4-yloxy]-propionic acid in
49% yield.
##STR00245##
Compound 42 was prepared in the same manner as Compound 41 using
42c instead of
(S)-1-{(S)-2-[(S)-2-(2-{4-[6-((R)-1-hydroxy-ethyl)-pyridin-2-yl]-phenoxy}-
-acetylamino)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-car-
boxylic acid 2,2,2-trichloro-ethyl ester in 8% yield. .sup.1H NMR
(300 MHz, CD.sub.3OD) 0.94 (d, J=6.7 Hz, 3H), 0.95 (d, J=6.5 Hz,
3H), 1.37 (d, J=7.1 Hz, 3H), 1.62 (d, J=6.5 Hz, 3H), 1.65-2.05 (m,
5H), 2.61-2.69 (m, 1H), 2.88-3.10 (m, 2H), 3.58-3.63 (m, 1H),
3.90-4.00 (m, 1H), 4.01 (d, J=8.5 Hz, 1H), 4.38-4.44 (m, 1H),
4.58-4.67 (m, 1H), 5.31 (q, J=7.1 Hz, 1H), 5.91 (q, J=6.5 Hz, 1H),
7.18 (d, J=7.6 Hz, 1H), 7.31 (app t, J=7.6 Hz, 1H), 7.47 (d, J=7.6
Hz, 1H), 7.58 (s, 1H), 7.77 (s, 1H), 7.95 (s, 1H). LCMS (m/z) 525.2
[M+H], Tr=4.51 min.
Example 43
##STR00246##
Potassium carbonate (663 mg, 4.80 mmol) and tetrakis
(triphenylphosphine)palladium(0) (139 mg, 0.12 mmol) were added to
a solution of (R)-1-(3-bromophenyl)-ethanol (483 mg, 2.40 mmol) and
4-(2-methoxy carbonylethyl) benzeneboronic acid (500 mg, 2.40 mmol)
in 1,2-dimethoxyethane (5 mL) in a 5 mL microwave vessel. The
vessel was sealed before being heated in the microwave for 20 min,
using fixed hold time, on high absorption at 100.degree. C. The
reaction mixture was filtered through a pad of Hyflo and the pad
was washed with ethyl acetate. The combined organics were then
concentrated and the resultant brown oil was purified by silica gel
chromatography using a stepwise gradient of iso-hexanes/ethyl
acetate 1:0 to 3:2 to afford the title compound (416 mg, 61%) as a
yellow oil.
##STR00247##
43a (416 mg, 1.46 mmol) was dissolved in a mixture of
tetrahydrofuran (8 mL) and water (2 mL) and the solution was cooled
using an ice bath. Lithium hydroxide monohydrate (175 mg, 2.92
mmol) was added and the solution was allowed to slowly warm to RT
overnight. The solution was acidified using 2 M hydrochloric acid
and then extracted with dichloromethane (2.times.20 mL). The
combined organics were then dried over anhydrous sodium sulfate,
filtered and concentrated to afford the title compound (250 mg,
89%) as a white solid.
##STR00248##
(S)-1-[(S)-2-((S)-2-Amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (669 mg, 1.55
mmol) and 43b (350 mg, 1.29 mmol) were dissolved in anhydrous
acetonitrile (10 mL) and cooled using an ice bath.
2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (688 mg, 1.81 mmol) and
N,N-diisopropylethylamine (899 .mu.L, 5.16 mmol) were then added
and the reaction was allowed to slowly warm to RT and left to stir
overnight. The solvent was then removed and the residue was
dissolved in ethyl acetate. The solution was then washed with water
(3.times.20 mL) and brine (20 mL), dried over anhydrous sodium
sulfate, filtered and concentrated and the resultant brown oil was
purified by silica gel chromatography using a stepwise gradient of
iso-hexanes/ethyl acetate 1:4 to 0:1 to afford the title compound
(314 mg, 36%) as a white solid.
##STR00249##
43c (314 mg, 0.459 mmol) was dissolved in tetrahydrofuran (10 mL).
Zinc powder (300 mg, 4.59 mmol) was added followed by ammonium
acetate (1 M in water, 3.21 mL, 3.21 mmol). The reaction was then
left to stir overnight. The reaction mixture was then filtered
through a pad of Hyflo. The pad was then washed with potassium
hydrogen sulfate solution and ethyl acetate. The biphasic mixture
was further acidified using 2 M hydrochloric acid until the
solution was approximately pH 1. The phases were separated and the
aqueous phase was extracted with ethyl acetate. The combined
organic phases were dried over anhydrous sodium sulfate, filtered
and concentrated to afford the title compound (212 mg, 84%) as a
yellow solid.
##STR00250##
43d (100 mg, 0.181 mmol) was dissolved in dichloromethane (181 mL),
under an atmosphere of nitrogen.
N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (142
mg, 0.724 mmol) and 4-dimethylaminopyridine (44 mg, 0.362 mmol)
were added and the reaction was left to stir overnight. The solvent
was removed and the residue was purified by silica gel
chromatography using 100% ethyl acetate to afford the title
compound (52 mg, 54%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta.0.92 (d, J=6.7 Hz, 6H), 1.24-1.29 (m, 3H),
1.60-1.68 (m, 5H), 1.85-1.98 (m, 2H), 2.37-2.49 (m, 1H), 2.58-2.78
(m, 1H), 2.83-2.93 (m, 1H), 3.16-3.28 (m, 1H), 3.45 (m, 3H),
4.03-4.11 (m, 1H), 4.45-4.54 (m, 1H), 5.06-5.16 (m, 1H), 5.92-6.00
(m, 1H), 6.25 (d, J=8.5 Hz, 1H), 7.19 (s, 1H), 7.22 (s, 2H), 7.39
(app t, J=7.6 Hz, 1H), 7.48-7.56 (m, 3H), 7.62 (br s, 1H). LCMS
(m/z) 535.3 [M+H], Tr=2.43 min.
Example 44
##STR00251##
Benzyl bromoacetate (2.07 mL, 13.1 mmol) and triphenylphosphine
(3.63 g, 13.8 mmol) were dissolved in toluene (60 mL) and left to
stir over 3 days. A white solid was collected by filtration and
washed with diethyl ether. This afforded the title compound (5.86
g, 93%) as a white solid.
##STR00252##
5-bromo-2-pyridine carboxaldehyde (125 mg, 0.666 mmol) and 44a (655
mg, 1.33 mmol) were dissolved in dichloromethane (5 mL).
1,1,3,3-Tetramethylguanidine (251 .mu.L, 2 mmol) was then added and
the reaction was left to stir for 3.5 h. The reaction was quenched
with saturated ammonium chloride solution and the phases were
separated. The organic phase was then washed with brine, dried over
anhydrous sodium sulfate, filtered and concentrated. The residue
was then purified by silica gel chromatography using a stepwise
gradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to afford the
title compound (186 mg, 88%) as a yellow solid.
##STR00253##
44b (186 mg, 0.585 mmol), (R)-1-[3-(4,4,5,5-tetramethyl-[1,3,2]
dioxaborolan-2-yl)-phenyl]-ethanol (145 mg, 0.585 mmol), cesium
carbonate (476 mg, 1.46 mmol) and tetrakis(triphenylphosphine)
palladium (0) (66 mg, 0.0585 mmol) were placed in a microwave
vessel. 1,2-Dimethoxyethane (2 mL) and water (0.5 mL) were added
and the vessel was sealed.
The reaction was heated in the microwave for 30 min, using fixed
hold time, on high absorption at 150.degree. C. The reaction
mixture was poured onto water and the resultant mixture was
extracted with ethyl acetate. The organics were then dried over
anhydrous sodium sulfate, filtered and concentrated. The residue
was then purified by silica gel chromatography using a stepwise
gradient of iso-hexanes/ethyl acetate 1:0 to 1:1 to afford the
title compound (150 mg, 77%) as an orange gum.
##STR00254##
44c (150 mg, 0.417 mmol) was dissolved in ethanol (5 mL).
Palladium(II) hydroxide (20% on carbon, wet, 40 mg) was then added
followed by ammonium formate (132 mg, 2.09 mmol). The reaction was
heated to reflux and left to stir for 30 min. The reaction was
allowed to cool to RT and was then filtered through a pad of Hyflo.
The pad was washed with ethanol and the combined organics were
concentrated to leave the title compound (105 mg, 93%) as an orange
gum.
##STR00255##
44d (105 mg, 0.387 mmol) and
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (200 mg,
0.464 mmol) were dissolved in acetonitrile (5 mL) and cooled using
an ice bath. 2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl
uronium hexafluorophosphate methanaminium (206 mg, 0.542 mmol) and
N,N-diisopropylethylamine (270 .mu.L, 1.55 mmol) were then added
and the reaction was allowed to slowly warm to RT and left to stir
overnight. The solvent was then removed and the residue was
dissolved in ethyl acetate. The solution was then washed with water
(3.times.20 mL) and brine (20 mL), dried over anhydrous magnesium
sulfate, filtered and concentrated and the residue was purified by
silica gel chromatography using a stepwise gradient of
iso-hexanes/ethyl acetate/acetone 3:1:0 to 0:7:3 to afford a brown
solid (110 mg). This was further purified by silica gel
chromatography using a stepwise gradient of ethyl acetate/acetone
1:0 to 4:1 to afford the title compound (100 mg, 38%) as a yellow
solid.
##STR00256##
44e (100 mg, 0.146 mmol) was dissolved in a mixture of
tetrahydrofuran (3 mL) and water (1 mL) and cooled using an ice
bath. Lithium hydroxide monohydrate (6 mg, 0.153 mmol) was added
and the reaction was stirred for 30 min. The solution was acidified
to pH 1 using 2 M hydrochloric acid and evaporated to dryness. The
residue was then purified by C18 chromatography using a stepwise
gradient of acetonitrile/water 0:1 to 1:4 to afford the title
compound (25 mg, 31%) as a white solid.
##STR00257##
44f (25 mg, 0.045 mmol) was dissolved in dichloromethane (45 mL).
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (35
mg, 0.148 mmol) and 4-dimethylaminopyridine (11 mg, 0.009 mmol)
were added and the reaction was left to stir for 4 h. The solvent
was removed and the residue was purified by silica gel
chromatography using a stepwise gradient of ethyl acetate/acetone
1:0 to 3:2 to leave a white solid (6 mg). This was then eluted
through an reverse phase HPLC system fitted with a Phenomenex
Gemini 10.mu. 110 A, 250.times.21.2 mm column using an isocratic
2:3 acetonitrile/water flow at 20 mL/min to afford the title
compound (1.3 mg, 7%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta.0.84-1.00 (m, 7H), 1.10 (d, J=6.9 Hz, 3H),
1.62-1.72 (m, 4H), 1.86-2.08 (m, 3H), 2.54-2.71 (m, 2H), 2.90-3.14
(m, 2H), 3.28-3.54 (m, 3H), 3.98 (app t, J=9.2 Hz, 1H), 4.43-4.52
(m, 1H), 5.07 (app t, J=8.0 Hz, 1H), 5.92-6.00 (m, 1H), 6.10 (d,
J=8.0 Hz, 1H), 6.37 (d, J=9.1 Hz, 1H), 7.20 (d, J=8.0 Hz, 1H),
7.26-7.31 (m, 1H), 7.41-7.59 (m, 3H), 7.75 (dd, J=8.0, 2.2 Hz, 1H),
8.78 (d, J=2.4 Hz, 1H). LCMS (m/z) 536.0 [M+H], Tr=1.47 min.
Example 45
##STR00258##
6-Bromo-3-pyridinecarboxaldehyde (500 mg, 2.69 mmol) and
benzyloxycarbonylmethyl-triphenyl-phosphonium bromide (2.64 g, 5.38
mmol) were dissolved in dichloromethane (15 mL).
1,1,3,3-Tetramethylguanidine (1.01 mL, 8.03 mmol) was then added
and the reaction was left to stir for 3.5 h. The reaction was
quenched with saturated ammonium chloride solution and the phases
were separated. The organic phase was then washed with brine, dried
over anhydrous sodium sulfate, filtered and concentrated. The
residue was then purified by silica gel chromatography using a
stepwise gradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to afford
the title compound (492 mg, 57%) as a white solid.
##STR00259##
45a (382 mg, 1.54 mmol),
(R)-1-[3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-ethanol
(490 mg, 1.54 mmol), cesium carbonate (1.06 g, 3.85 mmol) and
tetrakis(triphenylphosphine) palladium(0) (178 mg, 0.154 mmol) were
placed in a microwave vessel. 1,2-Dimethoxyethane (4 mL) and water
(1 mL) were added and the vessel was sealed. The reaction was
heated in the microwave for 50 min, using fixed hold time, on high
absorption at 150.degree. C. The reaction mixture was poured onto
water and the resultant mixture was extracted with ethyl acetate.
The organics were then dried over anhydrous sodium sulfate,
filtered and concentrated. The residue was then purified by silica
gel chromatography using a stepwise gradient of iso-hexanes/ethyl
acetate 1:0 to 1:1 to afford impure product. This was subjected to
a second round of purification using the same conditions to afford
the title compound (195 mg, 35%) as a yellow solid.
##STR00260##
45b (195 mg, 0.543 mmol) was dissolved in ethanol (5 mL).
Palladium(II) hydroxide (20% on carbon, wet, 40 mg) was then added
followed by ammonium formate (172 mg, 2.72 mmol). The reaction was
heated to reflux and left to stir for 30 min. The reaction was
allowed to cool to RT and was then filtered through a pad of Hyflo.
The pad was washed with ethanol and the combined organics were
concentrated to leave the title compound (150 mg, 100%) as a pale
yellow solid.
##STR00261##
45c (150 mg, 0.553 mmol) and
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (287 mg,
0.664 mmol) were dissolved in acetonitrile (5 mL) and cooled using
an ice bath. 2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl
uronium hexafluorophosphate methanaminium (294 mg, 0.774 mmol) and
N,N-diisopropylethylamine (385 .mu.L, 2.21 mmol) were then added
and the reaction was allowed to slowly warm to RT and left to stir
overnight. Ethyl acetate was then poured into the reaction mixture.
The resultant solution was then washed with water (3.times.20 mL)
and brine (20 mL), dried over anhydrous sodium sulfate, filtered
and concentrated and the residue was purified by silica gel
chromatography using a stepwise gradient of iso-hexanes/ethyl
acetate/acetone 3:1:0 to 0:7:3 to afford a solid (334 mg). This was
further purified by silica gel chromatography using a stepwise
gradient of ethyl acetate/acetone 1:0 to 4:1 to afford the title
compound (167 mg, 44%) as a solid.
##STR00262##
45d (160 mg, 0.234 mmol) was dissolved in a mixture of
tetrahydrofuran (4 mL) and water (1 mL) and cooled using an ice
bath. Lithium hydroxide monohydrate (6 mg, 0.153 mmol) was added
and the reaction was stirred for 30 min. The solution was
neutralized using 2 M hydrochloric acid and evaporated to dryness.
The residue was then purified by C18 chromatography using a
stepwise gradient of acetonitrile/water 0:1 to 1:4 to afford the
title compound (21 mg, 16%) as a white solid.
##STR00263##
45e (21 mg, 0.0379 mmol) was dissolved in dichloromethane (38 mL).
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (29
mg, 0.152 mmol) and 4-dimethylaminopyridine (9 mg, 0.0758 mmol)
were added and the reaction was left to stir for 4 h. The solvent
was removed and the residue was purified by silica gel
chromatography using 100% ethyl acetate. The resultant material was
then eluted through a reverse phase HPLC system fitted with a
Phenomenex Gemini 10.mu. 110 A, 250.times.21.2 mm column using an
isocratic 3:7 acetonitrile/water flow at 20 mL/min to afford the
title compound (1 mg, 5%). .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta.0.91 (d, J=6.5 Hz, 6H), 1.48 (d, J=7.1 Hz, 3H), 1.63-1.72
(m, 7H), 1.81-1.93 (m, 2H), 2.00-2.10 (m, 1H), 2.25-2.38 (m, 1H),
2.54-2.67 (m, 1H), 3.15-3.28 (m, 1H), 3.39-3.59 (m, 2H), 4.03 (app
t, J=9.6 Hz, 1H), 4.43-4.54 (m, 1H), 5.04-5.16 (m, 1H), 5.80 (d,
J=9.8 Hz, 1H), 5.94 (q, J=6.7 Hz, 1H), 7.22-7.27 (m, 1H), 7.43 (app
t, J=7.8 Hz, 1H), 7.55-7.57 (m, 1H), 7.69 (d, J=8.0 Hz, 1H), 7.92
(d, J=8.3 Hz, 1H), 8.19 (br s, 1H), 8.48 (d, J=2.2 Hz, 1H). LCMS
(m/z) 536.2 [M+H], Tr=1.61 min.
Example 46
##STR00264##
To Compound 21 (25 mg, 0.05 mmol) in ethyl acetate (5 mL) at RT was
added 10% palladium on carbon (20 mg). The system was purged with
hydrogen and stirred for 2 h. The reaction was filtered through
Celite and concentrated in vacuo. The residue was purified by
preparative TLC using ethyl acetate/acetone 5/1 to give the title
compound (2.2 mg, 9%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) 0.96 (d, J=6.7 Hz, 3H), 0.98 (d, J=6.7 Hz, 3H), 1.24
(d, J=7.1 Hz, 3H), 1.52 (d, J=6.9 Hz, 3H), 1.67 (d, J=6.7 Hz, 3H),
1.55-2.14 (m, 10H), 2.53-2.80 (m, 2H), 2.92-3.01 (m, 2H), 3.45-3.67
(m, 1H), 4.14-4.23 (m, 1H), 4.49-4.60 (m, 1H), 5.39-5.51 (m, 1H),
5.96 (d, J=9.4 Hz, 1H), 6.06 (q, J=6.7 Hz, 1H), 6.43 (d, J=8.5 Hz,
1H), 7.39 (dd, J=8.5, 1.1 Hz, 1H), 7.56 (s, 1H), 7.75 (s, 1H), 7.93
(d, J=8.5 Hz, 1H), 9.16 (s, 1H). LCMS (m/z)=552.3 [M+H], Tr=1.25
min.
Example 47
##STR00265##
To a solution of
(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-c-
arboxylic acid 2,2,2-trichloro-ethyl ester (320 mg, 0.74 mmol) in
dichloromethane (8 mL) was added trifluoroacetic acid (2 mL). The
reaction was stirred at RT for 105 min and was then concentrated in
vacuo. The resulting crude product was dissolved in ethyl acetate
(75 mL) and was washed with saturated aqueous sodium bicarbonate.
The aqueous phase was extracted with ethyl acetate (2.times.40 mL),
and the organics were dried over anhydrous sodium sulfate,
filtered, and concentrated to a crude residue that was used without
further purification. The residue from the previous step,
1-hydroxybenzotriazole (152 mg, 1.12 mmol) and
(S)-2-tert-butoxycarbonylamino-3-hydroxy-propionic acid (154 mg,
0.75 mmol) were dissolved in dichloromethane (6 mL).
N,N-Diisopropylethylamine (190 mg, 1.5 mmol) was added, and the
resulting solution was cooled in an ice water bath.
N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide (149 mg, 0.96 mmol)
was added dropwise over 15 s. The reaction was stirred for 18 h,
allowing the ice bath to slowly expire. The reaction mixture was
then diluted with ethyl acetate (50 mL) and was washed with
saturated aqueous sodium bicarbonate and brine. The organic phase
was dried over anhydrous sodium sulfate, filtered, and concentrated
to afford a crude residue that was purified by silica gel
chromatography (75 to 100% ethyl acetate in iso-hexanes) to afford
the title product (211 mg, 55%) as an oil.
##STR00266##
To a solution of 47a (64 mg, 0.12 mmol) in dichloromethane (1 mL)
was added 2,4,6-collidine (98 mg, 1.28 mmol).
tert-Butyldimethylsilyl trifluoromethanesulfonate (98 mg, 0.37
mmol) was added dropwise over 20 sec. The reaction mixture was
stirred for 15 h and was quenched with saturated aqueous sodium
bicarbonate (1 mL). The mixture was further diluted with ethyl
acetate, water, and 0.1 N aqueous hydrochloric acid to afford an
acidic aqueous layer. The phases were separated, and the organics
were dried over sodium sulfate, filtered, and concentrated in vacuo
to afford a crude residue that was used without further
purification. The crude product was dissolved in tetrahydrofuran (3
mL). Acetic acid (115 mg, 1.9 mmol) was added in one portion and
the resulting solution was stirred for 3.25 h. The reaction was
then diluted with ethyl acetate (25 mL) and washed with saturated
aqueous sodium bicarbonate (25 mL). The phases were separated, and
the aqueous phase was extracted with ethyl acetate (25 mL). The
combined organic phases were washed with brine (25 mL) and dried
over anhydrous sodium sulfate. The mixture was filtered, and the
filtrate was concentrated in vacuo to afford crude amine which was
used without further purification. The crude amine was dissolved in
N,N-dimethylformamide (1.5 mL). 2,4,6-Collidine (31 mg, 0.26 mmol)
and trans-3-pentenoic acid (15.3 mg, 0.152 mmol) were added, and
the resulting solution was cooled in an ice water bath.
(1-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeni-
um hexafluorophosphate (72.4 mg, 0.169 mmol) was added in one
portion, and the reaction was stirred for 30 min. The reaction was
then removed from the cold bath and warmed to ambient temperature.
After 15 h, the reaction was diluted with ethyl acetate (35 mL),
saturated aqueous sodium bicarbonate (20 mL), and brine (5 mL). The
phases were separated, and the organic layer was washed with 0.1 N
aqueous hydrochloric acid (25 mL) and then brine (5 mL). The acidic
aqueous layer was extracted with ethyl acetate (25 mL), and the
combined organic phases were then dried over anhydrous sodium
sulfate, filtered, and concentrated. The resulting crude residue
was purified by silica gel chromatography (50 to 80% ethyl acetate
in iso-hexanes) to afford the title compound (46.9 mg, 62% over 3
steps) as a white foam.
##STR00267##
To a solution of 47b (46.9 mg, 0.0761 mmol) in tetrahydrofuran (1.5
mL) was added water (0.30 mL), ammonium acetate (96 mg, 1.2 mmol),
and zinc powder (109 mg, 1.7 mmol). The reaction mixture was
stirred vigorously at RT for 17.5 h, at which time the temperature
was increased to 35.degree. C. After 25.5 h, additional zinc powder
(60 mg, 0.92 mmol) was added and the reaction temperature was
increased to 45.degree. C. After 39.5 h, the reaction mixture was
filtered through a pad of Celite washing with water and ethyl
acetate. The aqueous phase was acidified with 2 M aqueous
hydrochloric acid (15 mL), and the phases were separated. The
aqueous phase was extracted with ethyl acetate (2.times.20 mL), and
the combined organic phases were dried over sodium sulfate,
filtered, and concentrated to afford a white solid (34.9 mg, 95%)
that was used without further purification. The crude acid (0.072
mmol) was dissolved with (R)-1-(3-vinyl-isoquinolin-6-yl)-ethanol
(17.4 mg, 0.087 mmol) in dichloromethane (1.0 mL).
4-Dimethylaminopyridine (13.3 mg, 0.109 mmol) was added followed by
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (17
mg, 0.085 mmol). The reaction was stirred for 16.5 h, at which time
it was loaded directly onto a silica gel column. Elution with 60 to
100% ethyl acetate in iso-hexanes provided the title compound (20
mg, 43%) as an amorphous residue.
##STR00268##
A solution of 47c (19.6 mg, 0.0294 mmol) in toluene (9.4 mL) was
sparged with argon for 10 min with stirring. Hoveyda-Grubbs
2.sup.nd generation--catalyst (2.8 mg, 0.0045 mmol) was then added
as a solution in degassed toluene (0.45 mL), and the resulting
solution was heated to 105.degree. C. After 25 min, an additional
portion of Hoveyda-Grubbs 2.sup.nd generation catalyst (1.4 mg,
0.0022 mmol) was added. After 10 additional min, the reaction was
cooled to ambient temperature and was concentrated in vacuo to -4.5
mL. The solution was loaded directly onto a silica gel column which
was eluted with 80 to 100% ethyl acetate in iso-hexanes to afford
the title compound (6.9 mg, 38%) as an amorphous solid.
##STR00269##
In a polypropylene vial, 47d (4.7 mg, 0.0075 mmol) was dissolved in
tetrahydrofuran (0.90 mL) under argon and the resulting solution
was cooled in an ice water bath. HF.pyridine (0.10 mL) was added
dropwise. After 10 min, the reaction was quenched by addition to a
stirred mixture of ethyl acetate (15 mL) and saturated aqueous
sodium bicarbonate (15 mL). The phases were separated, and the
aqueous phase was extracted with ethyl acetate (3.times.30 mL). The
combined organic phases were dried over sodium sulfate, filtered
and concentrated to afford a crude residue. In a polypropylene vial
under argon, the aforementioned residue was dissolved in
tetrahydrofuran (0.90 mL) and the resulting solution was cooled in
an ice water bath. HF.pyridine (0.10 mL) was added dropwise, and
the reaction mixture was removed from the cold bath. After 45 min,
the reaction was worked up as described above to afford a crude
residue. Purification by reverse-phase HPLC (5 to 100% acetonitrile
in water, +0.1% trifluoroacetic acid) provided the title compound
(1.6 mg, 34%) as an amorphous white solid. .sup.1H NMR (400 MHz,
CD.sub.3OD) 9.54 (s, 1H), 8.38 (d, J=8.7 Hz, 1H), 8.29 (s, 1H),
8.17 (s, 1H), 7.87 (d, J=8.5 Hz, 1H), 6.90-6.75 (m, 2H), 6.18 (q,
J=6.9 Hz, 1H), 5.84-5.74 (m, 1H), 4.71-4.64 (m, 1H), 4.50-4.42 (m,
1H), 3.88-3.81 (m, 2H), 3.78 (dd, J=11.3, 6.4 Hz, 1H), 3.52 (dd,
J=12.9, 6.2 Hz, 1H), 3.52 (dd, J=12.9, 6.2 Hz, 1H), 2.81-2.70 (m,
1H), 2.11-2.04 (m, 1H), 1.99-1.91 (m, 1H), 1.85-1.69 (m, 2H), 1.73
(d, J=6.7 Hz, 3H), 1.60 (d, J=7.1 Hz, 3H). LCMS (m/z) 510.2 [M+H],
Tr=2.19 min.
Example 48
##STR00270##
To a solution of
(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-c-
arboxylic acid 2,2,2-trichloro-ethyl ester (320 mg, 0.74 mmol) in
dichloromethane (8.8 mL) was added trifluoroacetic acid (2.2 mL).
The reaction was stirred at RT for 45 min and was concentrated in
vacuo. The resulting crude product was twice dissolved in and
concentrated from anhydrous toluene (10 mL). The resulting crude
residue was used without further purification. The crude residue
(ca. 0.74 mmol), 1-hydroxybenzotriazole (153.8 mg, 1.1 mmol), and
(2S,3R)-2-tert-butoxycarbonylamino-3-hydroxy-butyric acid (161.2
mg, 0.735 mmol) were dissolved in dichloromethane (4 mL).
N,N-Diisopropylethylamine (192 mg, 1.5 mmol) was added, and the
resulting solution was cooled in an ice water bath.
N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide (132 mg, 0.85 mmol)
was added dropwise over 15 s. The reaction was removed from the
cold bath and was stirred for 21.5 h. The reaction mixture was then
diluted with ethyl acetate (50 mL) and washed with half-saturated
aqueous sodium bicarbonate (30 mL). The organic phase was washed
with brine, dried over anhydrous sodium sulfate, filtered, and
concentrated to afford a crude residue that was purified by silica
gel chromatography eluting with 65 to 90% ethyl acetate in
iso-hexanes. Impure fractions were repurified by silica gel
chromatography and collated with the first product to provide the
title compound (230 mg, 58%).
##STR00271##
To a solution of 48a (62.3 mg, 0.12 mmol) in dichloromethane (1 mL)
was added 2,4,6-collidine (146 mg, 1.21 mmol).
tert-Butyldimethylsilyl trifluoromethanesulfonate (94 mg, 0.36
mmol) was added dropwise over 20 s. The reaction mixture was
stirred for 15 h and was quenched with saturated aqueous sodium
bicarbonate (1 mL). The mixture was further diluted with ethyl
acetate (2 mL) and brine (1 mL). The phases were separated, and the
aqueous phase was extracted with ethyl acetate (4.times.1.5 mL).
The combined organic phases were dried over sodium sulfate,
filtered, and concentrated in vacuo to afford a crude residue that
was used without further purification. The crude product was
dissolved in tetrahydrofuran (3 mL). Acetic acid (104 mg, 1.6 mmol)
was added in one portion and the resulting solution was stirred for
3.5 h. The reaction was then diluted with ethyl acetate (30 mL),
saturated aqueous sodium bicarbonate (20 mL) and brine (10 mL). The
phases were separated, and the aqueous phase was extracted with
ethyl acetate (20 mL). The combined organic phases were dried over
anhydrous sodium sulfate. The mixture was filtered, and the
filtrate was concentrated in vacuo to afford crude intermediate
amine which was used without further purification. The crude amine
was dissolved in N,N-dimethylformamide (1.5 mL). 2,4,6-Collidine
(28 mg, 0.23 mmol) and trans-3-pentenoic acid (14.3 mg, 0.143 mmol)
were added, and the resulting solution was cooled in an ice water
bath.
(1-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeni-
um hexafluorophosphate (69 mg, 0.16 mmol) was added in one portion,
and the reaction was stirred for 30 min. The reaction was then
removed from the cold bath and warmed to ambient temperature. After
15 h, the reaction was diluted with ethyl acetate (25 mL), 0.1 N
aqueous hydrochloric acid (30 mL), and brine (5 mL). The phases
were separated, and the acidic aqueous layer was extracted with
ethyl acetate (25 mL). The combined organic phases were washed with
saturated aqueous sodium bicarbonate (25 mL), with brine (5 mL),
dried over anhydrous sodium sulfate, filtered, and concentrated.
The resulting crude residue was purified by silica gel
chromatography (50 to 75% ethyl acetate in iso-hexanes) to afford
the title compound (74 mg, quantitative yield over 3 steps) as a
white foam.
##STR00272##
To a solution of 48b (74 mg, 0.117 mmol) in tetrahydrofuran (2.3
mL) was added water (0.47 mL), ammonium acetate (138 mg, 1.8 mmol),
and zinc powder (164 mg, 2.5 mmol). The reaction mixture was
stirred vigorously at RT for 17.5 h, at which time the temperature
was increased to 35.degree. C. After 25.5 h, additional zinc powder
(85 mg, 1.3 mmol) was added and the reaction temperature was
increased to 45.degree. C. After 39.5 total hours, the reaction
mixture was filtered through a pad of Celite, washing with water
and ethyl acetate. The aqueous phase was acidified with 2 N aqueous
hydrochloric acid (15 mL), and the phases were separated. The
aqueous phase was extracted with ethyl acetate (2.times.20 mL), and
the combined organic phases were dried over anhydrous sodium
sulfate, filtered, and concentrated to afford an amorphous white
solid (50 mg, 86%) that was used without further purification. The
crude product (0.10 mmol) was dissolved along with
(R)-1-(3-vinyl-isoquinolin-6-yl)-ethanol (23.9 mg, 0.12 mmol) in
dichloromethane (1.0 mL). 4-Dimethylaminopyridine (15 mg, 0.12
mmol) was added followed by
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (17
mg, 0.019 mmol). The reaction was stirred for 16 h, at which time
it was loaded directly onto a silica gel column. Elution with 50 to
85, and then to 100% ethyl acetate in iso-hexanes provided the
title compound (40 mg, 58%) as a solid.
##STR00273##
A solution of 48c (26 mg, 0.037 mmol) in toluene (12 mL) was
sparged with argon for several min with stirring. Hoveyda-Grubbs
2.sup.nd generation catalyst (3.4 mg, 0.0054 mmol) was then added
as a solution in degassed toluene (0.40 mL), and the resulting
solution was heated to 105.degree. C. After 20 min, an additional
portion of Hoveyda-Grubbs 2.sup.nd generation catalyst (1.7 mg,
0.0027 mmol) was added as a solution in toluene (0.20 mL). After 40
additional min, the reaction was cooled to RT and was concentrated
in vacuo to -3 mL. The solution was loaded directly onto a silica
gel column, which was eluted with 70 to 100% ethyl acetate in
iso-hexanes to afford the title compound (10 mg, 42%) as an
amorphous solid.
##STR00274##
In a polypropylene vial under argon atmosphere, 48d (10 mg, 0.016
mmol) was dissolved in tetrahydrofuran (900 .mu.L). HF.pyridine
(.about.70% as HF, 100 .mu.L) was added dropwise via syringe, and
the resulting solution was stirred for 130 min. An additional
aliquot of HF.pyridine (100 .mu.L) was then added via syringe, and
the resulting solution was stirred for an additional 80 min. The
reaction mixture was quenched by its careful addition to a stirred
mixture of ethyl acetate (50 mL) and saturated aqueous sodium
bicarbonate (50 mL). The phases were separated, and the aqueous
phase was extracted with ethyl acetate (2.times.30 mL). The
combined organic phases were dried over anhydrous sodium sulfate,
filtered, and concentrated to afford a crude residue that was
purified by reverse phase HPLC (C18, 15 to 100% acetonitrile/water,
0.1% trifluoroacetic acid). Impure fractions from this run were
re-purified in the same manner to afford the title compound as the
trifluoroacetic acid salt (5.0 mg, 49%) as an amorphous white solid
following lyophilization. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.9.55 (s, 1H), 8.38 (d, J=8.6 Hz, 1H), 8.22 (s, 1H), 8.15 (s,
1H), 7.87 (dd, J=8.7, 1.4 Hz, 1H), 6.86-6.75 (m, 2H), 6.15 (q,
J=6.6 Hz, 1H), 5.80-5.70 (m, 1H), 4.47 (d, J=5.6 Hz, 1H), 4.45-4.38
(m, 1H), 4.09-4.01 (m, 1H), 3.83-3.78 (m, 1H), 3.55 (dd, J=13.5,
5.6 Hz, 1H), 3.11 (dd, J=13.2, 4.7 Hz, 1H), 2.78-2.67 (m, 1H),
2.09-1.99 (m, 1H), 1.96-1.89 (m, 1H), 1.82-1.64 (m, 5H), 1.60-1.54
(m, 3H), 1.25 (d, J=6.3 Hz, 3H). LCMS (m/z) 524.6 [M+H], Tr=2.15
min.
Example 49
##STR00275##
A solution of
(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-c-
arboxylic acid 2,2,2-trichloro-ethyl ester (865 mg, 2 mmol) in
dichloromethane (20 mL) was cooled in an ice water bath.
Trimethylsilyl trifluoromethanesulfonate (667 mg, 3 mmol) was added
dropwise at 0.degree. C. under argon, and the resulting solution
was stirred at RT for 30 min. The reaction mixture was evaporated
to dryness and the resulting crude residue was dissolved in
anhydrous acetonitrile (25 mL) under argon. The reaction mixture
was stirred at 0.degree. C.,
(S)-4-azido-2-(tert-butoxycarbonylamino)butanoic acid
dicyclohexylamine salt (936 mg, 2.2 mmol) and
N,N-diisopropylethylamine (1034 mg, 8 mmol) were added followed by
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (1065 mg, 2.8 mmol). The reaction
mixture was stirred at RT for 48 h. The solvent was evaporated, the
residue was dissolved in ethyl acetate (100 mL) and the solution
was washed with 20% water solution of citric acid (2.times.100 mL),
water (100 mL) and brine (100 mL), dried over magnesium sulfate,
filtered and evaporated. The residue was purified by silica gel
chromatography (gradient from 0-40% ethyl acetate+methanol (4/1) in
iso-hexanes) to afford the title compound (783 mg, 70%) as a white
solid after evaporation. R.sub.f=0.40, iso-hexanes/ethyl
acetate/methanol (6/4/1).
##STR00276##
Under argon, (R)-1-(3-chloro-isoquinolin-6-yl)-ethanol (880 mg,
4.23 mmol),
(E)-2,2-dimethyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)--
but-3-enoic acid methyl ester (1.24 g, 4.88 mmol),
PdCl.sub.2(PCy.sub.2(p-NMe.sub.2Ph)).sub.2(bis[(dicyclohexyl)(4-dimethyla-
minophenyl)phosphine]palladium(II)chloride) 173 mg, 0.21 mmol) and
potassium phosphate tribasic (2.64 g, 12.4 mmol) were dissolved in
cyclopentyl methyl ether (11.9 mL) and water (5.1 mL). The
resulting biphasic mixture was vigorously stirred at 90.degree. C.
for 3.5 h, at which time the reaction was cooled to ambient
temperature and was diluted with ethyl acetate (50 mL) and water
(40 mL). The phases were separated, and the aqueous phase was
extracted with ethyl acetate (2.times.50 mL). The combined organic
phases were dried over anhydrous magnesium sulfate, filtered, and
concentrated to afford a crude residue that was purified by silica
gel chromatography (25 to 60% ethyl acetate in iso-hexanes) to
afford the title compound (1.07 g, 85%) as a yellow oil.
##STR00277##
To a solution of 49b (600 mg, 2 mmol) in tetrahydrofuran (8 mL) was
added methanol (4 mL), water (4 mL) and lithium hydroxide (96 mg, 4
mmol). The resulting mixture was stirred at RT for 10 h and
quenched with 1 M hydrochloric acid (4.2 mL, 4.2 mmol). The
resulting solution was concentrated to a crude residue which was
co-distilled twice with tetrahydrofuran (20 mL), twice with
anhydrous acetonitrile (20 mL) and twice with anhydrous toluene (20
mL). The resulting white solid was dried under high vacuum
overnight and it was used without further purification (735 mg,
quantitative yield).
##STR00278##
A solution of 49a (169 mg, 0.302 mmol) in dichloromethane (10 mL)
was cooled in an ice water bath. Trimethylsilyl
trifluoromethanesulfonate (101 mg, 0.455 mmol) was added dropwise
at 0.degree. C. under argon, and the resulting solution was stirred
at RT for 30 min. The reaction mixture was evaporated to dryness
and the resulting crude residue was dissolved in anhydrous
acetonitrile (20 mL) under argon. Reaction mixture was stirred at
0.degree. C.,
(E)-4-[6-((R)-1-hydroxy-ethyl)-isoquinolin-3-yl]-2,2-dimethyl-but-3-enoic
acid (123 mg, 0.333 mmol) and N,N-diisopropylethylamine (151 mg,
1.220 mmol) were added followed by
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (161 mg, 0.423 mmol). The
reaction mixture was stirred at RT for 2 days. The solvent was
evaporated, the residue was dissolved in ethyl acetate (50 mL) and
the solution was washed with 20% water solution of citric acid
(2.times.50 mL), water (50 mL) and brine (50 mL), dried over
magnesium sulfate, filtered and evaporated. The residue was
purified by silica gel chromatography (gradient from 0-40% ethyl
acetate and methanol mixture (4/1) in iso-hexanes) to afford the
title compound (198 mg, 90%) as a white solid after evaporation.
R.sub.f=0.18, iso-hexanes/ethyl acetate/methanol (6/4/1).
##STR00279##
To a solution of 49d (170 mg, 0.23 mmol) in tetrahydrofuran (2 mL)
was added methanol (1 mL), water (1 mL) and lithium hydroxide
hydrate (7 mg, 0.28 mmol). The mixture was stirred for 2 h at
ambient temperature and was quenched with aqueous 1 M hydrochloric
acid (0.30 mL, 0.30 mmol). The resulting solution was concentrated
to a crude residue which was co-evaporated twice with
tetrahydrofuran (5 mL), twice with anhydrous acetonitrile (5 mL)
and twice with anhydrous toluene (5 mL). The resulting white solid
was dried under high vacuum overnight and used without further
purification (151 mg, quantitative yield). Into oven-dried, argon
purged flask were placed 2-methyl-6-nitrobenzoic anhydride (317 mg,
0.92 mmol), 4-dimethylaminopyridine (337 mg, 2.76 mmol) and
anhydrous 1,2-dichloroethane (300 mL). The resulting solution was
heated at 50.degree. C., and the crude seco-acid was added dropwise
by syringe as a solution in dry N,N-dimethylformamide (5 mL) over
12 h. An additional portion of dry N,N-dimethylformamide (2.times.1
ml) was used to complete the quantitative transfer. After stirring
for additional 2 h at 50.degree. C., the reaction mixture was
transferred to a separatory funnel and washed with water (200 mL,
10 mL of brine was added to support the separation). The aqueous
phase was extracted with dichloromethane (100 mL). Combined organic
extracts were washed with brine (100 mL) and dried over magnesium
sulfate, filtered and concentrated under reduced pressure. The
resulting residue was dissolved in ethyl acetate (250 mL) and was
washed with water (300 mL, 10 mL of brine was added to support the
separation). The aqueous phase was extracted with ethyl acetate
(150 mL). Combined organic extracts were washed with water (200 mL,
10 mL of brine was added to support the separation). Resulting
aqueous phase was extracted with ethyl acetate (150 mL). Combined
organic extracts were washed with brine (150 mL) and dried over
magnesium sulfate, filtered and concentrated under reduced
pressure. The residue was purified by silica gel chromatography
(gradient from 0-40% ethyl acetate and methanol mixture (4/1) in
iso-hexanes) to afford the title compound (76 mg, 57%) as a white
solid after evaporation. R.sub.f=0.51, 10% methanol in
dichloromethane. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.9.03 (s,
1H), 7.95 (d, J=8.5 Hz, 1H), 7.81 (s, 1H), 7.49 (s, 1H), 7.46 (dd,
J=8.5, 1.6 Hz, 1H), 6.49 (d, J=16.1 Hz, 1H), 6.40 (d, J=16.1 Hz,
1H), 5.95 (q, J=6.6 Hz, 1H), 5.51 (q, J=7.2 Hz, 1H), 4.67 (dd,
J=8.6, 6.3 Hz, 1H), 4.29 (m, 1H), 3.68 (dd, J=11.2, 2.7 Hz, 1H),
3.28 (td, J=6.8, 3.7 Hz, 2H), 2.67-2.56 (m, 1H), 1.88 (m, 2H),
1.85-1.60 (m, 4H), 1.57 (d, J=6.7 Hz, 3H), 1.54 (d, J=7.3 Hz, 3H),
1.42 (s, 3H), 1.27 (s, 3H). LCMS (m/z) 557.3 ([M+H], Tr=3.15
min.
Example 50
##STR00280##
Into an oven-dried, argon purged flask, Compound 49 (20 mg, 0.034
mmol), copper(I) iodide (1 mg, 0.005 mmol) and propiolic acid (5
mg, 0.070 mmol) were added. The flask was sealed with septa and
repurged with argon three times. Anhydrous N,N-dimethylformamide (5
mL) was added and the reaction mixture was repurged with argon
three times. This reaction mixture was heated at 70.degree. C. for
2 days. After evaporation of the solvent under reduced pressure,
the crude residue was dissolved in ethyl acetate (10 mL) and
filtered through Celite and the filter pad was washed with ethyl
acetate (10 mL). After concentration under reduced pressure, the
residue was purified by silica gel chromatography (gradient from
0-40% ethyl acetate and methanol mixture (4/1) in iso-hexanes) to
afford the title compound (7 mg, 34%) as a white solid.
R.sub.f=0.36, 5% methanol in dichloromethane. .sup.1H NMR (400 MHz,
CD.sub.3OD): .delta.9.05 (s, 1H), 8.11 (br s, 1H), 8.00 (br s, 1H),
7.95 (d, J=8.8 Hz, 1H), 7.82 (s, 1H), 7.66 (s, 1H), 7.48 (d, J=10.0
Hz, 1H), 6.50 (d, J=16.2 Hz, 1H), 6.42 (d, J=16.2 Hz, 1H), 5.97 (m,
1H), 5.56 (m, 1H), 4.60 (m, 1H), 4.29 (m, 1H), 3.66 (m, 1H), 3.21
(m, 2H), 2.69-2.47 (m, 1H), 1.89 (m, 2H), 1.87-1.55 (m, 4H), 1.53
(d, J=6.7 Hz, 3H), 1.50 (d, J=7.1 Hz, 3H), 1.40 (s, 3H), 1.28 (s,
3H). LCMS (m/z) 603.1 [M+H]' Tr=2.62 min.
Example 51
##STR00281##
A solution of
(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-c-
arboxylic acid 2,2,2-trichloro-ethyl ester (865 mg, 2 mmol) in
dichloromethane (20 mL) was cooled in an ice water bath.
Trimethylsilyl trifluoromethanesulfonate (667 mg, 3 mmol) was added
dropwise at 0.degree. C. under argon, and the resulting solution
was stirred at RT for 30 min. The reaction mixture was evaporated
to dryness and the resulting crude residue was dissolved in
anhydrous acetonitrile (25 mL) under argon. Reaction mixture was
stirred at 0.degree. C.,
(S)-2-(tert-butoxycarbonylamino)pent-4-ynoic acid (469 mg, 2.2
mmol, source: Matrix Scientific, Catalog Number 041479) and
N,N-diisopropylethylamine (1034 mg, 8 mmol) were added followed by
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (1065 mg, 2.8 mmol). Reaction
mixture was stirred at RT for 48 h. The solvent was evaporated, the
residue was dissolved in ethyl acetate (100 mL) and the solution
was washed with 20% water solution of citric acid (2.times.100 mL),
water (100 mL) and brine (100 mL), dried over magnesium sulfate,
filtered and evaporated. The residue was purified by silica gel
chromatography (gradient from 0-40% ethyl acetate and methanol
(4/1) in iso-hexanes) to afford the title compound (738 mg, 70%) as
a white solid after evaporation. R.sub.f=0.30, iso-hexanes/ethyl
acetate/methanol (6/4/1).
##STR00282##
A solution of 51a (145 mg, 0.275 mmol) in dichloromethane (10 mL)
was cooled in an ice water bath. Trimethylsilyl
trifluoromethanesulfonate (92 mg, 0.414 mmol) was added dropwise at
0.degree. C. under argon, and the resulting solution was stirred at
RT for 30 min. The reaction mixture was evaporated to dryness and
the resulting crude residue was dissolved in anhydrous acetonitrile
(20 mL) under argon. The reaction mixture was stirred at 0.degree.
C.,
(E)-4-[6-((R)-1-hydroxy-ethyl)-isoquinolin-3-yl]-2,2-dimethyl-but-3-enoic
acid (112 mg, 0.303 mmol) and N,N-diisopropylethylamine (137 mg,
1.101 mmol) were added followed by
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (147 mg, 0.385 mmol). The
reaction mixture was stirred at RT for 2 days. The solvent was
evaporated, the residue was dissolved in ethyl acetate (50 mL) and
the solution was washed with 20% water solution of citric acid
(2.times.50 mL), water (50 mL) and brine (50 mL), dried over
magnesium sulfate, filtered and evaporated. The residue was
purified by silica gel chromatography (gradient from 0-40% ethyl
acetate and methanol (4/1) in iso-hexanes) to afford the title
compound (163 mg, 85%) as a white solid after evaporation.
R.sub.f=0.29, iso-hexanes/ethyl acetate/methanol (6/4/1).
##STR00283##
To a solution of 51b (125 mg, 0.18 mmol) in tetrahydrofuran (2 mL)
was added methanol (1 mL), water (1 mL) and lithium hydroxide
hydrate (5.2 mg, 0.22 mmol). The mixture was stirred for 2 h at
ambient temperature and was quenched with aqueous 1 M hydrochloric
acid (0.25 mL, 0.25 mmol). The resulting solution was concentrated
to a crude residue which was co-evaporated twice with
tetrahydrofuran (5 mL), twice with anhydrous acetonitrile (5 mL)
and twice with anhydrous toluene (5 mL). The resulting white solid
was dried under high vacuum overnight and it was used without
further purification (113 mg, quantitative yield). Into an
oven-dried, argon purged flask were placed 2-methyl-6-nitrobenzoic
anhydride (248 mg, 0.72 mmol), 4-dimethylaminopyridine (264 mg,
2.16 mmol) and anhydrous 1,2-dichloroethane (200 mL). The resulting
solution was heated at 50.degree. C., and the crude seco-acid was
added dropwise via syringe as a solution in anhydrous
N,N-dimethylformamide (5 mL) over 12 h. An additional portion of
anhydrous N,N-dimethylformamide (2.times.1 ml) was used to complete
the quantitative transfer. After stirring for additional 2 h at
50.degree. C., the reaction mixture was transferred to a separatory
funnel and washed with water (200 mL, 10 mL of brine was added to
support the separation). The aqueous phase was extracted with
dichloromethane (100 mL). Combined organic extracts were washed
with brine (100 mL) and dried over magnesium sulfate, filtered and
concentrated under reduced pressure. The resulting residue was
dissolved in ethyl acetate (250 mL) and was washed with water (300
mL, 10 mL of brine was added to support the separation). The
aqueous phase was extracted with ethyl acetate (150 mL). Combined
organic extracts were washed with water (200 mL, 10 mL of brine was
added to support the separation). Resulting aqueous phase was
extracted with ethyl acetate (150 mL). Combined organic extracts
were washed with brine (150 mL) and dried over magnesium sulfate,
filtered and concentrated under reduced pressure. The residue was
purified by silica gel chromatography (gradient from 0-40% ethyl
acetate+methanol (4/1) in iso-hexanes) to afford the title compound
(56 mg, 57%) as a white solid after evaporation. R.sub.f=0.53, 10%
methanol in dichloromethane. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.9.03 (s, 1H), 7.94 (d, J=8.5 Hz, 1H), 7.79 (s, 1H), 7.54 (s,
1H), 7.45 (d, J=8.5 Hz, 1H), 6.52 (d, J=16.1 Hz, 1H), 6.40 (d,
J=16.1 Hz, 1H), 5.95 (q, J=6.6 Hz, 1H), 5.52 (q, J=7.2 Hz, 1H),
4.70 (dd, J=7.7, 6.7 Hz, 1H), 4.30 (m, 1H), 4.00 (q, J=7.2 Hz, 1H),
3.72-3.64 (m, 1H), 2.63 (m, 1H), 2.55-2.40 (m, 2H), 1.91 (s, 1H),
1.88 (m, 1H), 1.80 (m, 1H), 1.64 (m, 1H), 1.57 (d, J=6.7 Hz, 3H),
1.53 (d, J=7.2 Hz, 3H), 1.42 (s, 3H), 1.27 (s, 3H). LCMS (m/z)
546.2 [M+H]' Tr=3.04 min.
Example 52
##STR00284##
A solution of Compound 51 (10 mg, 0.018 mmol) in a mixture of ethyl
acetate (4 mL) and tetrahydrofuran (4 mL) containing 5% palladium
on calcium carbonate poisoned with lead--Lindlar catalyst (10 mg)
was hydrogenated at RT and pressure of hydrogen for 3 h. The
reaction mixture was filtered through Celite and the filter pad was
washed with tetrahydrofuran (10 mL). The filtrate was evaporated to
afford the title compound (10 mg, quantitative yield) as a white
solid. R.sub.f=0.19, iso-hexanes/ethyl acetate/methanol (6/4/1).
.sup.1H NMR (400 MHz, CD.sub.3OD): .delta.9.04 (s, 1H), 7.95 (d,
J=8.4 Hz, 1H), 7.82 (s, 1H), 7.54 (s, 1H), 7.46 (d, J=8.3 Hz, 1H),
6.50 (d, J=16.1 Hz, 1H), 6.39 (d, J=16.1 Hz, 1H), 5.95 (q, J=6.4
Hz, 1H), 5.76-5.62 (m, 1H), 5.57 (q, J=7.0 Hz, 1H), 5.02 (d, J=17.0
Hz, 1H), 4.96 (d, J=10.1 Hz, 1H), 4.61-4.54 (m, 1H), 4.29 (m, 1H),
3.68n (m, 1H), 2.61 (m, 1H), 2.42-2.34 (m, 1H), 2.30-2.20 (m, 1H),
1.90 (m, 1H), 1.81 (m, 1H), 1.68-1.60 (m, 2H), 1.57 (d, J=6.6 Hz,
3H), 1.53 (d, J=7.2 Hz, 3H), 1.39 (s, 3H), 1.26 (s, 3H). LCMS (m/z)
548.3 [M+H]' Tr=2.85 min.
Example 53
##STR00285##
To a solution of
(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-c-
arboxylic acid 2,2,2-trichloro-ethyl ester (300 mg, 0.69 mmol) in
dichloromethane (1.84 mL) was slowly added trifluoroacetic acid
(460 .mu.L, 6.00 mmol) at 0.degree. C. under an argon atmosphere.
After 3 h, the reaction mixture was concentrated under reduced
pressure. The crude residue was dissolved in N,N-dimethylformamide
(3.45 mL) and (S)-tert-Butoxycarbonylamino-cyclohexyl-acetic acid
(195 mg, 0.760 mmol),
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (289 mg, 0.760 mmol), and
N,N-diisopropylethylamine (180 .mu.L, 1.04 mmol) were sequentially
added at 23.degree. C. under an argon atmosphere. After 18 h, the
reaction mixture was diluted with ethyl acetate (300 mL), and the
resulting mixture was washed with brine (4.times.100 mL), was dried
over anhydrous sodium sulfate, and was concentrated under reduced
pressure. The crude residue was purified by silica gel
chromatography (12 g Combiflash HP Gold Column, 0-100% ethyl
acetate/iso-hexanes gradient) to afford the title compound (380 mg,
96%) as a colorless oil.
##STR00286##
To a solution of 53a (220 mg, 0.385 mmol) in dichloromethane (1.92
mL) was added trimethylsilyl trifluoromethanesulfonate (128 mg,
0.587 mmol) at 0.degree. C. under an argon atmosphere. After 1.5 h,
the reaction mixture was concentrated under reduced pressure. The
resulting residue was diluted with acetonitrile (1.92 mL) and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (146 mg, 0.385 mmol),
N,N-diisopropylethylamine (267 .mu.L, 1.54 mmol), and
(E)-pent-3-enoic acid (39.4 .mu.L, 0.385 mmol) were sequentially
added at 23.degree. C. under an argon atmosphere. After 20 h, the
reaction mixture was diluted with dichloromethane (40 mL) and the
resulting mixture was washed with saturated aqueous sodium
bicarbonate solution (40 mL) and with brine (2.times.40 mL). The
organic layer was separated, was dried over anhydrous sodium
sulfate, and was concentrated under reduced pressure. The crude
residue was purified by silica gel flash column chromatography (12
g Combiflash HP Gold Column, 0-100% ethyl acetate/iso-hexanes
gradient) to afford the title compound (180 mg, 84%) as a colorless
oil. R.sub.f=0.75 (ethyl acetate) I.sub.2/silica stain.
##STR00287##
To a solution of 53b (180 mg, 0.320 mmol) in tetrahydrofuran (5.3
mL) was added zinc powder (418 mg, 6.40 mmol) followed by a
solution of ammonium acetate (370 mg, 4.80 mmol) in water (3.5 mL)
at 23.degree. C. under an argon atmosphere. After 15 h, the
reaction mixture was warmed to 45.degree. C. After 2 h, the
reaction mixture was allowed to cool to RT and was filtered through
a pad of Celite washing with water (10 mL) and ethyl acetate (10
mL). The filtrate layers were split and the aqueous layer was
diluted with brine (80 mL) and was acidified to pH 1 with 12 N
aqueous hydrogen chloride solution. The aqueous layer was extracted
with ethyl acetate (3.times.100 mL), and the combined organic
extracts were dried over anhydrous sodium sulfate, and were
concentrated under reduced pressure. Residual acetic acid was
removed azeotropically via addition of toluene (5 mL) followed by
concentration under reduced pressure (3.times.) to afford the title
compound (74.5 mg, 55%) as a white solid.
##STR00288##
To a solution of 53c (74.5 mg, 0.176 mmol) and
(R)-1-(3-vinyl-isoquinolin-6-yl)-ethanol (42.1 mg, 0.212 mmol) in
dichloromethane (3.5 mL) were added
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (47.2
mg, 0.246 mmol) and 4-dimethylaminopyridine (11 mg, 88 .mu.mol) at
23.degree. C. under an argon atmosphere. After 18 h, the reaction
mixture was purified directly by silica gel chromatography (12 g
Combiflash HP Gold Column, 0-100% ethyl acetate/iso-hexanes
gradient) to afford the title compound (67.6 mg, 64%) as a white
solid.
##STR00289##
To a solution of 53d (25 mg, 41 .mu.mol) in toluene (8.2 mL) was
added the Hoveyda-Grubbs 2.sup.nd Generation catalyst (2.5 mg, 4.1
.mu.mol) at 23.degree. C. under an argon atmosphere, and the
resulting mixture was heated to 110.degree. C. After 2 h, the
reaction mixture was quenched with ethyl vinyl ether (300 .mu.L)
and the resulting mixture was allowed to cool to 23.degree. C. The
reaction mixture was concentrated under reduced pressure and the
crude residue was purified by silica gel chromatography (12 g
Combiflash HP Gold Column, 0-100% ethyl acetate/iso-hexanes
gradient). The fractions containing the desired product were
combined and were repurified by preparatory HPLC to afford the
title compound (1.7 mg, 7%) as a white powder as a trifluoroacetic
acid salt. R.sub.f=0.40 (ethyl acetate) UV. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta.9.13 (s, 1H), 8.53 (br s, 1H), 8.07 (d, J=7.6
Hz, 1H), 8.05 (d, J=8.5 Hz, 1H), 7.83 (s, 1H), 7.55 (d, J=8.3 Hz,
1H), 7.44 (s, 1H), 6.62 (d, J=16.3 Hz, 1H), 6.55-6.45 (m, 1H), 6.04
(q, J=6.5 Hz, 1H), 5.50 (q, J=6.9 Hz, 1H), 4.40 (d, J=14.1 Hz, 1H),
4.33 (d, J=9.7 Hz, 1H), 3.78 (d, J=11.0 Hz, 1H), 3.36 (app dd,
J=14.7, 6.7 Hz, 1H), 2.99 (dd, J=14.5, 4.9 Hz, 1H), 2.90-2.66 (m,
4H), 2.02-1.83 (m, 4H), 1.82-1.70 (m, 3H), 1.68 (d, J=6.7 Hz, 3H),
1.65 (d, J=7.3 Hz, 3H), 1.33-1.16 (m, 4H), 1.16-0.94 (m, 2H). HPLC
Tr=t.sub.R (min), 3.091 (Synergi 4u hydro-RP, 50.times.4.60 mm 4
micron column, 7 min, 2 ml/min, 5-100% acetonitrile/water, 0.05%
trifluoroacetic acid modifier gradient). LCMS (m/z) 562.3 [M+H],
Tr=2.17 min.
Example 54
##STR00290##
To a solution of
(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-c-
arboxylic acid 2,2,2-trichloro-ethyl ester (300 mg, 0.69 mmol) in
dichloromethane (1.84 mL) was slowly added trifluoroacetic acid
(460 .mu.L, 6.00 mmol) at 0.degree. C. under an argon atmosphere.
After 2 h, the reaction mixture was concentrated under reduced
pressure. The crude residue was dissolved in N,N-dimethylformamide
(3.45 mL) and (S)-2-tert-Butoxycarbonylamino-3-phenyl-propionic
acid (201 mg, 0.760 mmol),
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (289 mg, 0.760 mmol), and
N,N-diisopropylethylamine (180 .mu.L, 1.04 mmol) were sequentially
added at 23.degree. C. under an argon atmosphere. After 22 h, the
reaction mixture was diluted with ethyl acetate (300 mL), and the
resulting mixture was washed with brine (3.times.200 mL), was dried
over anhydrous sodium sulfate, and was concentrated under reduced
pressure. The crude residue was purified by silica gel
chromatography (24 g Combiflash HP Gold Column, 0-100% ethyl
acetate/iso-hexanes gradient) to afford the title compound (319 mg,
96%) as a colorless oil. R.sub.f=0.75 (ethyl acetate)
I.sub.2/silica stain.
##STR00291##
To a solution of 54a (414 mg, 0.716 mmol) in dichloromethane (3.58
mL) was added trimethylsilyl trifluoromethanesulfonate (238.7 mg,
1.07 mmol) at 0.degree. C. under an argon atmosphere. After 1 h,
the reaction mixture was concentrated under reduced pressure. The
resulting residue was diluted with acetonitrile (3.58 mL) and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (272 mg, 0.716 mmol),
N,N-diisopropylethylamine (498 .mu.L, 2.86 mmol), and
(E)-pent-3-enoic acid (73.3 .mu.L, 0.716 mmol) were sequentially
added at 23.degree. C. under an argon atmosphere. After 17 h, the
reaction mixture was diluted with dichloromethane (50 mL) and the
resulting mixture was washed with saturated aqueous sodium
bicarbonate solution (50 mL). The organic layer was separated, was
dried over anhydrous sodium sulfate, and was concentrated under
reduced pressure. The crude residue was purified by silica gel
chromatography to afford the title compound (386 mg, 96%) as white
solid.
##STR00292##
To a solution of 54b (443 mg, 0.789 mmol) in tetrahydrofuran (13.2
mL) was added zinc powder (1.03 g, 15.8 mmol) followed by a
solution of ammonium acetate (912 mg, 11.8 mmol) in water (8.77 mL)
at 23.degree. C. under an argon atmosphere. After 17 h, the
reaction mixture was warmed to 45.degree. C. After 2 h, the
reaction mixture was allowed to cool to RT and was filtered through
a pad of Celite washing with water (10 mL) and ethyl acetate (10
mL). The filtrate layers were split and the aqueous layer was
diluted with brine (20 mL) and was acidified to pH 2 with 12 N
aqueous hydrogen chloride solution. The aqueous layer was extracted
with dichloromethane (3.times.100 mL), and the combined organic
extracts were dried over anhydrous sodium sulfate, and were
concentrated under reduced pressure. Residual acetic acid was
removed azeotropically via addition of toluene (5 mL) followed by
concentration under reduced pressure (3.times.) to afford the title
compound (276.2 mg, 81%) as a white solid.
##STR00293##
To a solution of 54c (275 mg, 0.640 mmol) and
(R)-1-(3-vinyl-isoquinolin-6-yl)-ethanol (153 mg, 0.770 mmol) in
dichloromethane (3.2 mL) were added
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (172
mg, 0.90 mmol) and 4-dimethylaminopyridine (39 mg, 32 .mu.mol) at
23.degree. C. under an argon atmosphere. After 23 h, the reaction
mixture was purified directly by silica gel chromatography (24 g
Combiflash HP Gold Column, 0-100% ethyl acetate/iso-hexanes
gradient) to afford the title compound (203 mg, 52%) as a white
solid.
##STR00294##
To a solution of 54d (170 mg, 278 .mu.mol) in chlorobenzene (56 mL)
was added the Hoveyda-Grubbs 2.sup.nd Generation catalyst (8.7 mg,
14.0 .mu.mol) at 23.degree. C. under an argon atmosphere, and the
resulting mixture was heated to 110.degree. C. After 3 h, the
reaction mixture was quenched with ethyl vinyl ether (300 .mu.L)
and the resulting mixture was allowed to cool to 23.degree. C. The
reaction mixture was concentrated under reduced pressure and the
crude residue was purified by silica gel chromatography (12 g
Combiflash HP Gold Column, 0-100% ethyl acetate/iso-hexanes
gradient) to afford the title compound (18.9 mg, 12%) as a tan
solid. R.sub.f=0.25 (ethyl acetate) UV. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta.9.12 (s, 1H), 8.05 (d, J=8.7 Hz, 1H), 7.84 (s,
1H), 7.56 (d, J=8.5 Hz, 1H), 7.53 (s, 1H), 7.32-7.12 (m, 5H), 6.66
(d, J=16.5 Hz, 1H), 6.48 (dt, J=12.8, 5.6 Hz, 1H), 6.05 (q, J=6.4
Hz, 1H), 5.53 (q, J=6.7 Hz, 1H), 4.73 (d, J=12.2 Hz, 1H), 4.43 (d,
J=11.9 Hz, 1H), 3.80 (app t, J=10.2 Hz, 1H), 3.30-3.22 (m, 1H),
3.09 (dd, J=14.5, 4.9 Hz, 1H), 2.96-2.69 (m, 3H), 2.04-1.87 (m,
2H), 1.82-1.56 (m, 2H), 1.68 (d, J=6.5 Hz, 3H), 1.64 (d, J=6.9 Hz,
3H). HPLC Tr=3.060 min. LCMS (m/z) 570.5 [M+H], Tr=2.14 min.
Example 55
##STR00295##
To a solution of
(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-c-
arboxylic acid 2,2,2-trichloro-ethyl ester (500 mg, 0.1.16 mmol) in
dichloromethane (5.8 mL) was added trimethylsilyl
trifluoromethanesulfonate (386 mg, 1.74 mmol) at 0.degree. C. under
an argon atmosphere. After 1 h, the reaction mixture was
concentrated under reduced pressure. The resulting residue was
diluted with acetonitrile (5.8 mL) and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (485 mg, 1.28 mmol),
N,N-diisopropylethylamine (302 .mu.L, 1.74 mmol),
N-tert-butoxycarbnyl-3-(3-pyridyl)-L-alanine (337 mg, 1.27 mmol)
were sequentially added at 23.degree. C. under an argon atmosphere.
After 20 h, the reaction mixture was concentrated under reduced
pressure, and the crude residue was purified by silica gel
chromatography (40 g Combiflash HP Gold Column, 0-100% ethyl
acetate/iso-hexanes gradient) to afford the title compound (570 mg,
85%) as light yellow oil.
##STR00296##
To a solution of 55a (570 mg, 0.984 mmol) in dichloromethane (5.8
mL) was added trimethylsilyl trifluoromethanesulfonate (386 mg,
1.74 mmol) at 0.degree. C. under an argon atmosphere. After 1 h,
the reaction mixture was concentrated under reduced pressure to
afford the title compound (478 mg) as light yellow oil which was
used without further purification.
##STR00297##
To a solution of 55b (120 mg, 0.250 mmol) in acetonitrile (1.25 mL)
were sequentially added
(E)-4-[6-((R)-1-hydroxy-ethyl)-isoquinolin-3-yl]-2,2-dimethyl-but-3-enoic
acid (75 mg, 0.25 mmol),
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (95 mg, 0.25 mmol), and
N,N-diisopropylethylamine (173 .mu.L, 1.00 mmol) at 23.degree. C.
under an argon atmosphere. N,N-Dimethylformamide (100 .mu.L) was
then added to promote solubility of the reagents. After 23 h, the
reaction mixture was diluted with saturated aqueous sodium
bicarbonate solution (25 mL) and brine (25 mL) and the resulting
mixture was extracted with dichloromethane (2.times.25 mL). The
combined organic extracts were dried over anhydrous sodium sulfate
and were concentrated under reduced pressure. The crude residue was
purified by silica gel chromatography to afford the title compound
(76 mg, 41%) as a colorless solid.
##STR00298##
To a solution of 55c (76 mg, 0.10 mmol) in tetrahydrofuran (0.3 mL)
and water (0.2 mL) was added lithium hydroxide hydrate (2.4 mg,
0.10 mmol) at 23.degree. C. under an argon atmosphere. After 2 h,
the reaction mixture was concentrated under reduced pressure to
afford the title compound (61 mg, 99%) as a white solid lithium
carboxylate salt.
##STR00299##
To a solution of 55d (61 mg, 0.10 mmol) in dichloromethane (50 mL)
were added benzotriazol-1-yl-oxytripyrrolidinophosphonium
hexafluorophosphate (208 mg, 400 .mu.mol) and
4-dimethylaminopyridine (366 mg, 3.00 mmol) at 23.degree. C. under
an argon atmosphere. After 17 h, the reaction mixture was
concentrated under reduced pressure and the crude residue was
purified by silica gel chromatography (4 g Combiflash HP Gold
Column, 0-20% methanol/dichloromethane gradient) to afford the
title compound (1.2 mg, 2%) as a colorless solid. R.sub.f=0.40 (10%
methanol in dichloromethane) UV. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.9.03 (s, 1H), 8.31 (d, J=1.6 Hz, 1H), 8.27 (dd, J=4.9, 1.5
Hz, 1H), 7.95 (d, J=8.5 Hz, 1H), 7.82 (s, 1H), 7.64 (d, J=8.0 Hz,
1H), 7.53 (s, 1H), 7.46 (dd, J=8.5, 1.5 Hz, 1H), 7.24 (dd, J=7.9,
4.9 Hz, 1H), 6.49 (d, J=16.1 Hz, 1H), 6.37 (d, J=16.1 Hz, 1H), 5.95
(q, J=6.5 Hz, 1H), 5.57 (q, J=7.3 Hz, 1H), 4.65 (d, J=12.0 Hz, 1H),
4.32 (br d, J=12.1 Hz, 1H), 3.83 (d, J=11.8 Hz, 1H), 3.73-3.65 (m,
1H), 3.00 (dd, J=14.2, 5.0 Hz, 1H), 2.83 (dd, J=13.9, 9.7 Hz, 1H),
2.64 (br t, J=11.2 Hz, 1H), 1.95-1.73 (m, 2H), 1.69-1.59 (m, 1H),
1.57 (d, J=6.7 Hz, 3H), 1.54 (d, J=7.2 Hz, 3H), 1.33 (s, 3H), 1.16
(s, 3H). HPLC Tr=4.491 min
Example 56
##STR00300##
To a solution of
(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-c-
arboxylic acid 2,2,2-trichloro-ethyl ester (791 mg, 1.82 mmol) in
dichloromethane (10.0 mL) was slowly added trimethylsilyl
trifluoromethanesulfonate (483 .mu.L, 2.73 mmol) at 0.degree. C.
under an argon atmosphere. After 45 min, the reaction mixture was
concentrated under reduced pressure. The resulting residue was
diluted with dichloromethane (10.0 mL) and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (761 mg, 2.00 mmol),
N,N-diisopropylethylamine (1.26 mL, 7.28 mmol), and
(S)-2-(tert-butoxycarbonylamino)-4-phenylbutanoic acid (Fluka, 560
mg, 2.00 mmol) were sequentially added at 23.degree. C. under an
argon atmosphere. After 18 h, the reaction mixture was concentrated
under reduced pressure. The residue was pre-absorbed on silica and
purified by silica gel chromatography (40 g Isco Rf Gold Column,
0-100% ethyl acetate/iso-hexanes gradient) to afford the title
compound (799 mg, 74%) as a colorless oil.
##STR00301##
To a solution of 56a (799 mg, 1.34 mmol) in dichloromethane (10.0
mL) was added trimethylsilyl trifluoromethanesulfonate (356 .mu.L,
2.01 mmol) at 0.degree. C. under an argon atmosphere. After 30 min,
the reaction mixture was concentrated under reduced pressure. The
resulting residue was diluted with acetonitrile (6.0 mL) and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (560 mg, 1.47 mmol),
N,N-diisopropylethylamine (932 .mu.L, 5.36 mmol), and
(E)-pent-3-enoic acid (150 .mu.L, 1.47 mmol) were sequentially
added at 23.degree. C. under an argon atmosphere. After 19 h, the
reaction mixture was concentrated under reduced pressure. The
residue was pre-absorbed on silica and purified by silica gel
chromatography to afford the title compound (598 mg, 78%) as a
colorless oil.
##STR00302##
To a solution of 56b (167 mg, 0.29 mmol) in tetrahydrofuran (3 mL),
methanol (1 mL) and water (1 mL) was added lithium hydroxide
hydrate (7.6 mg, 0.32 mmol) at 23.degree. C. After 15 min, the
reaction mixture was concentrated to dryness under reduced
pressure. The residue was dissolved in water (25 mL) and washed
with ethyl acetate. The organic layer was extracted with saturated
aqueous sodium bicarbonate solution (25 mL). The combined aqueous
layers were acidified with 1 M aqueous hydrochloric acid solution
to pH .about.2 and extracted with ethyl acetate (3.times.25 mL),
and the combined organic extracts were dried over anhydrous
magnesium sulfate, and were concentrated under reduced pressure to
afford the title compound (129 mg, 100%) as a white solid.
##STR00303##
To a solution of 56c (129 mg, 0.29 mmol) and
(R)-1-(3-vinyl-isoquinolin-6-yl)-ethanol (64 mg, 0.31 mmol) in
dichloromethane (3.0 mL) was added
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (89
mg, 0.46 mmol) and 4-dimethylaminopyridine (18 mg, 0.14 mmol) at
23.degree. C. under an argon atmosphere. After 16 h, the reaction
mixture was purified directly by silica gel chromatography to
afford the title compound (87 mg, 48%) as a white solid.
##STR00304##
To a solution of 56d (84 mg, 0.13 mmol) in chlorobenezene (27 mL)
was added the Hoveyda-Grubbs 2.sup.nd Generation catalyst (8 mg, 13
.mu.mol) at 23.degree. C. under an argon atmosphere, and the
resulting mixture was heated to 110.degree. C. After 2 h, the
Hoveyda-Grubbs 2.sup.nd Generation catalyst (7.3 mg, 11 .mu.mol)
was added under argon at 110.degree. C. The Hoveyda-Grubbs 2.sup.nd
Generation Catalyst (5 mg, 8 .mu.mol) was then added in the
interval of 30 min for three times at which point the reaction was
complete. The reaction mixture was quenched with ethyl vinyl ether
(1.0 .mu.L) and the resulting mixture was allowed to cool to
23.degree. C. The reaction mixture was concentrated under reduced
pressure and the crude residue was purified by silica gel
chromatography (24 g Isco Rf Gold Column, 0-100% ethyl
acetate/iso-hexanes gradient for 10 min and then 100% ethyl acetate
for 25 min) to afford the title compound (9.1 mg, 12%) as a pale
brown solid. R.sub.f=0.20 (ethyl acetate). .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta.9.00 (s, 1H), 7.93 (d, J=8.5 Hz, 1H), 7.71 (s,
1H), 7.45 (dd, J=8.5, 1.5 Hz, 1H), 7.36 (s, 1H), 7.20-7.08 (m, 4H),
7.08-7.00 (m, 1H), 6.55 (d, J=15.9 Hz, 1H), 6.40 (ddd, J=15.9, 7.0,
5.1 Hz, 1H), 5.95 (q, J=6.5 Hz, 1H), 5.40 (q, J=7.2 Hz, 1H), 4.60
(d, J=12.1 Hz, 1H), 4.53 (dd, J=8.2, 6.9 Hz, 1H), 4.30 (d, J=12.5
Hz, 1H), 3.70-3.63 (m, 1H), 3.31-3.23 (m, 1H), 2.93 (ddd, J=14.0,
5.1, 1.6 Hz, 1H), 2.69-2.52 (m, 3H), 2.01-1.73 (m, 4H), 1.71-1.54
(m, 2H), 1.59 (d, J=6.7 Hz, 3H), 1.46 (d, J=7.3 Hz, 3H). HPLC
Tr=5.108 min. LCMS (m/z) 584.3 [M+H], Tr=2.12 min.
Example 57
##STR00305##
A solution of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methylbutyrylamino)-propio-
nyl]-hexahydropyridazine-3-carboxylic acid 2,2,2-trichloroethyl
ester (1.61 g, 3.03 mmol) in dichloromethane (31 mL) was cooled in
an ice water bath under argon. Trimethylsilyl
trifluoromethanesulfonate (1.23 g, 5.5 mmol) was added dropwise,
and the resulting solution was stirred for 2 h. The reaction was
quenched with N,N-diisopropylethylamine (1.2 g, 9.2 mmol) and
methanol (8.5 mL). The mixture was concentrated in vacuo and was
redissolved and concentrated from toluene (2.times.25 mL). The
resulting crude residue containing
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (1.05 g) was
used without further purification.
##STR00306##
To a solution of
(E)-4-[6-((R)-1-hydroxy-ethyl)-isoquinolin-3-yl]-2,2-dimethyl-but-3-enoic
acid methyl ester (1.05 g, 3.51 mmol) in tetrahydrofuran (8 mL) was
added methanol (4 mL), water (4 mL), and lithium hydroxide hydrate
(297 mg, 7.08 mmol). The resulting mixture was stirred for 6 h and
was quenched with 1 M aqueous hydrochloric acid (7.2 mL, 7.2 mmol).
The resulting solution was concentrated to a crude residue that was
redissolved and concentrated from anhydrous methanol (50 mL)
followed by toluene (50 mL). The resulting yellow solid (1.3 g,
100%)
(E)-4-[6-((R)-1-hydroxy-ethyl)-isoquinolin-3-yl]-2,2-dimethyl-but-3-enoic
acid was used without further purification. A portion of crude
(E)-4-[6-((R)-1-hydroxy-ethyl)-isoquinolin-3-yl]-2,2-dimethyl-but-3-enoic
acid (1.05 g, 2.84 mmol) was dissolved in N,N-dimethylformamide (12
mL) under argon. N,N-diisopropylethylamine (1.85 g, 14.3 mmol) was
added followed by 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl
uronium hexafluorophosphate methanaminium (1.42 g, 3.73 mmol). The
resulting mixture was stirred for 3 min, at which time crude 57a
was added as a solution in N,N-dimethylformamide (8.5 mL), washing
with additional N,N-dimethylformamide (2.times.5 mL). The reaction
was stirred for 40 min and was then diluted with ethyl acetate (200
mL) and water (300 mL). The phases were separated, and the aqueous
phase was extracted with ethyl acetate (200 mL). The combined
organic phases were washed with water (150 mL), and the second
aqueous layer was extracted with ethyl acetate (100 mL). The
combined organics were dried over anhydrous sodium sulfate,
filtered, and concentrated. The crude residue was purified by
silica gel chromatography (35 to 60% acetone in iso-hexanes,
continuous gradient) to afford the title compound (1.51 g, 71% over
2 steps) as a colorless oil. R.sub.F 0.5 (50% acetone in
iso-hexanes).
##STR00307##
To a solution of 57b (1.50 g, 2.15 mmol) in tetrahydrofuran (20 mL)
was added methanol (10 mL), water (10 mL), and lithium hydroxide
hydrate (365 mg, 8.7 mmol). The mixture was stirred for 75 min at
ambient temperature and was then quenched with aqueous 1 M
hydrochloric acid (8.8 mL, 8.8 mmol). The resulting solution was
concentrated in vacuo, and the crude product was twice dissolved
and concentrated from methanol (40 mL) and suspended and
concentrated from acetonitrile (6.times.30 mL) to afford 1.71 g of
a colorless solid that was used without further purification. Under
argon, 2-methyl-6-nitrobenzoic anhydride (1.85 g, 5.37 mmol) and
4-dimethylaminopyridine (1.97 g, 16.1 mmol) were dissolved in
1,2-dichloroethane (700 mL). The resulting solution was heated to
50.degree. C., and the crude seco-acid was added dropwise via
syringe as a solution in N,N-dimethylformamide (22 mL) over 6 h. An
additional wash with N,N-dimethylformamide (1.5 mL) was then added
in the same manner over 15 min. After stirring an additional 1.25
h, the reaction mixture was concentrated to a final volume of 200
mL in vacuo. The solution was washed with water (250 mL), and the
aqueous phase was extracted with dichloromethane (150 mL). The
organic phase was dried over anhydrous magnesium sulfate, filtered,
and concentrated. The resulting residue was dissolved in ethyl
acetate (200 mL) and was washed with water (150 mL). The aqueous
phase was extracted with ethyl acetate (150 mL). The combined
organics were washed with water (100 mL), and the second aqueous
phase was extracted with ethyl acetate (100 mL). The combined
organics were dried over anhydrous magnesium sulfate, filtered and
concentrated. The crude residue was purified by silica gel
chromatography (35 to 65% acetone in iso-hexanes continuous
gradient) to afford 445 mg of the pure title compound as an
amorphous white solid along with 389 mg of impure product. The
impure fractions were purified by recrystallization from
acetone:iso-hexanes to afford an additional 173.5 mg of pure title
compound (total: 618.5 mg, 52%). R.sub.F 0.5 (50% acetone in
iso-hexanes). .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.9.13 (s,
1H), 8.04 (d, J=8.5 Hz, 1H), 7.89 (s, 1H), 7.55 (dd, J=8.5, 1.6 Hz,
1H), 7.53 (s, 1H), 6.73-6.34 (m, 2H), 6.03 (q, J=6.6 Hz, 1H), 5.63
(q, J=7.2 Hz, 1H), 4.67 (d, J=12.2 Hz, 1H), 4.47-4.21 (m, 2H),
3.81-3.73 (m, 1H), 2.76-2.64 (m, 1H), 2.08-1.81 (m, 3H), 1.80-1.58
(m, 8H), 1.51 (s, 3H), 1.35 (s, 3H), 0.98 (d, J=6.7 Hz, 3H), 0.93
(d, J=6.7 Hz, 3H). LCMS (m/z) 550.2 [M+H], Tr=2.74 min.
Example 58
##STR00308##
A solution of N,N-diisopropylamine (2.51 g, 24.8 mmol) in
tetrahydrofuran (150 mL) under argon was cooled in an ice water
bath. A solution of n-butyllithium in hexanes (2.5 M, 9.7 mL, 24
mmol) was added dropwise over 2 min, and the resulting solution was
stirred for 15 additional minutes. The solution was then cooled to
-78.degree. C. via CO.sub.2(s):acetone bath, and ethyl
1-acetylcyclopropanecarboxylate (3.47 g, 22.2 mmol) was added
dropwise over 2 min. The solution was stirred for an additional 20
min, and N-Phenyl-bis(trifluoromethanesulfonimide) (8.4 g, 23.5
mmol) was added as a solution in tetrahydrofuran (24 mL) via canula
over 5 min, washing with additional portions of tetrahydrofuran
(2.times.5 mL). The resulting solution was removed from the cold
bath. After an additional 30 min, the reaction mixture was
concentrated in vacuo and was diluted with diethyl ether (200 mL).
The organic phase was washed with 1 M aqueous sodium hydroxide
(1.times.100 mL, 1.times.30 mL), dried over anhydrous magnesium
sulfate, filtered, and concentrated to afford 6.6 g of a crude oil
that was used without further purification. A solution of the crude
material from the previous step in anhydrous pyridine (11 mL) was
heated to 90.degree. C. After 16.5 h, the reaction mixture was
diluted with diethyl ether (200 mL) and 3 M hydrochloric acid (100
mL) and the phases were separated. The organic phase was washed
with 3 M hydrochloric acid (50 mL) and 1 M sodium hydroxide (50
mL), dried over magnesium sulfate, filtered, and concentrated to
afford 2.2 g of a crude liquid that was used without further
purification. To an argon-flushed vessel containing zirconocene
dichloride (410 mg, 1.6 mmol) was added a solution of the crude
product from the previous step (2.2 g, ca. 16 mmol) and
pinacolborane (3.1 g, 24 mmol) in dichloromethane (8 mL). After 116
h, the stirred reaction mixture was diluted with ethyl acetate (50
mL) and was quenched by dropwise addition of water. The mixture was
further diluted with water (50 mL), and the phases were separated.
The aqueous phase was extracted with ethyl acetate (2.times.50 mL).
The combined organic phases were dried over magnesium sulfate,
filtered, and concentrated to a crude residue that was purified by
silica gel chromatography (5 to 20% ethyl acetate in iso-hexanes,
continuous gradient) to afford the title compound (1.26 g, 21% over
3 steps) as an oil that crystallized on standing at -15.degree.
C.
##STR00309##
To a round-bottomed flask was added
(R)-1-(3-chloroisoquinolin-6-yl)ethanol (204 mg, 0.982 mmol), 58a
(314 mg, 1.18 mmol), PdCl.sub.2(PCy.sub.2(p-NMe.sub.2Ph)).sub.2
(bis[(dicyclohexyl)(4-dimethylaminophenyl)phosphine]palladium(II)
chloride) (40 mg, 0.049 mmol) and potassium phosphate tribasic (680
mg, 3.2 mmol). The vessel was sealed with a septum cap and was
flushed with argon. Cyclopentyl methyl ether (2.8 mL) and water
(1.2 mL) were added, and the resulting biphasic mixture was
vigorously stirred in an oil bath pre-heated to 90.degree. C. After
6.75 h, the reaction was cooled to ambient temperature and was
diluted with ethyl acetate (50 mL) and water (40 mL). The phases
were separated, and the aqueous phase was extracted with ethyl
acetate (50 mL). The combined organic phases were dried over
magnesium sulfate, filtered, and concentrated to afford a crude
residue that was purified by silica gel chromatography (35 to 60%
ethyl acetate in iso-hexanes) to afford the title compound (266 mg,
85%).
##STR00310##
To a solution of 58b (266 mg, 0.854 mmol) in tetrahydrofuran (2 mL)
was added methanol (1 mL), water (1 mL), and lithium hydroxide
hydrate (70.8 mg, 1.69 mmol). The resulting mixture was stirred for
3 h and was then quenched with 1 M aqueous hydrochloric acid (1.8
mL, 1.8 mmol). Volatiles were removed in vacuo, and the resulting
solid was suspended in toluene. Volatiles were removed in vacuo,
and the resulting yellow solid
1-{(E)-2-[6-((R)-1-hydroxy-ethyl)-isoquinolin-3-yl]-vinyl}-cyclopropaneca-
rboxylic acid was used without further purification. A solution of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methylbutyrylamino)-propio-
nyl]-hexahydropyridazine-3-carboxylic acid 2,2,2-trichloroethyl
ester (486 mg, 0.914 mmol) in dichloromethane (9.4 mL) was cooled
in an ice water bath under argon. Trimethylsilyl
trifluoromethanesulfonate (370 mg, 1.7 mmol) was added dropwise,
and the resulting solution was stirred for 4 h. The reaction was
quenched with N,N-diisopropylethylamine (360 mg, 2.7 mmol) and
methanol (2.5 mL). The mixture was concentrated in vacuo and the
resulting residue was redissolved and concentrated from toluene
(2.times.15 mL). The resulting crude amine was used without further
purification. To a solution of crude
1-{(E)-2-[6-((R)-1-hydroxy-ethyl)-isoquinolin-3-yl]-vinyl}-cyclopropaneca-
rboxylic acid (ca. 0.854 mmol) in N,N-dimethylformamide (3.5 mL)
under argon was added N,N-diisopropylethylamine (560 mg, 4.3 mmol)
and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (427 mg, 1.12 mmol). The
resulting mixture was stirred for 2 min, at which time crude
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester from the
previous step was added as a solution in N,N-dimethylformamide (2.5
mL), washing with two additional portions of N,N-dimethylformamide
(1.5 mL each). The reaction was stirred for 45 min and was diluted
with ethyl acetate (100 mL) and water (150 mL). The phases were
separated, and the aqueous phase was extracted with ethyl acetate
(100 mL). The combined organic phases were washed with water (75
mL), and the second aqueous layer was extracted with ethyl acetate
(75 mL). The combined organics were dried over anhydrous magnesium
sulfate, filtered, and concentrated. The crude residue was purified
by silica gel chromatography (35 to 60% acetone in iso-hexanes,
continuous gradient) to afford title compound (467 mg, 78% over 2
steps) as a colorless oil.
##STR00311##
To a solution of 58c (467 mg, 0.67 mmol) in tetrahydrofuran (6 mL)
was added methanol (3 mL), water (3 mL), and lithium hydroxide
hydrate (113 mg, 2.69 mmol). The mixture was stirred for 1.75 h at
ambient temperature and was quenched with aqueous 1 M hydrochloric
acid (2.8 mL, 2.8 mmol). The resulting solution was concentrated in
vacuo, and the crude product was suspended and concentrated from
acetonitrile (5.times.20 mL) to afford 530 mg of a pale yellow
solid that was used without further purification. Under argon,
2-methyl-6-nitrobenzoic anhydride (283 mg, 0.82 mmol) and
4-dimethylaminopyridine (307 mg, 2.51 mmol) were dissolved in
1,2-dichloroethane (100 mL). The resulting solution was heated to
50.degree. C., and a portion of the crude seco-acid (260 mg, ca.
0.33 mmol) was added dropwise via syringe as a solution in
N,N-dimethylformamide (3.5 mL) and 1,2-dichloroethane (10 mL) over
6 h. An additional wash of N,N-dimethylformamide (1 mL) was then
added in the same manner. After stirring an additional 1.25 h, the
reaction mixture was concentrated to -35 mL in vacuo. The solution
was diluted with ethyl acetate (100 mL) and washed with water (100
mL). The aqueous phase was extracted with ethyl acetate (75 mL),
and the combined organic phases were washed with water (50 mL). The
second aqueous phase was extracted with ethyl acetate (50 mL), and
the combined organics were dried over magnesium sulfate, filtered
and concentrated. The crude residue was purified by silica gel
chromatography (35 to 60 and then to 100% acetone in iso-hexanes)
to afford 101 mg of impure product containing the title compound.
This material was purified by silica gel chromatography (0 to 5%
methanol in ethyl acetate) followed by reverse-phase HPLC (5 to
100% acetonitrile/water+0.1% trifluoroacetic acid) to afford the
title compound as its trifluoroacetic acid salt (37 mg, 17% over 2
steps). .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.9.55 (s, 1H), 8.37
(d, J=8.6 Hz, 1H), 8.06 (s, 1H), 7.89 (s, 1H), 7.85 (d, J=8.6 Hz,
1H), 6.69 (d, J=16.4 Hz, 1H), 6.54 (d, J=16.4 Hz, 1H), 6.11 (q,
J=6.7 Hz, 1H), 5.75-5.67 (m, 1H), 4.46-4.36 (m, 1H), 4.36-4.28 (m,
1H), 3.90-3.78 (m, 1H), 2.80-2.68 (m, 1H), 2.05-1.86 (m, 3H),
1.80-1.68 (m, 5H), 1.66-1.57 (m, 5H), 1.40-1.30 (m, 1H), 1.30-1.21
(m, 1H), 1.03-0.93 (m, 4H), 0.90 (d, J=6.7 Hz, 3H). LCMS (m/z)
548.4 [M+H], Tr=2.79 min.
Example 59
##STR00312##
Into an oven dried, argon purged flask were placed oil-free
potassium hydride (from 1740 mg of ca. 30% dispersion in mineral
oil, ca. 13 mmol), anhydrous tetrahydrofuran (10 mL) and hexane (1
mL). The flask was repurged with argon and trichloroethylene (900
.mu.L, 1.32 g, 10 mmol) was added followed by dry methanol (10
.mu.L, 7.9 mg, 0.25 mmol). This mixture was stirred at RT for two
h. After this time, hexane (10 mL) was added and the resulting
solution was immediately used in the subsequent step.
##STR00313##
Into an oven dried, argon purged flask tetrahydrofuran (50 mL) was
added and the solution was cooled with an ice bath. A 1.8 M
solution of lithium diisopropylamide (7.2 mL, 13 mmol) in
tetrahydrofuran/heptane/ethylbenzene was added. The resulting
solution was cooled to -78.degree. C., and treated dropwise with
methyl tetrahydro-2H-pyran-4-carboxylate (1.20 mL. 1.30 g, 9 mmol)
followed by hexamethylphosphoramide (1.56 mL, 1.61 g, 9 mmol). The
resulting solution was warmed to 0.degree. C., stirred for 20 min.,
cooled to -78.degree. C., and treated dropwise with pre-cooled
(0.degree. C.) solution of 1,2-dichloro-ethyne (ca. 10 mmol). The
reaction mixture was stirred at -78.degree. C. for 30 min and then
allowed to warm to RT. After 4 h at RT, the reaction mixture was
poured into crushed ice and extracted with diethyl ether (200 mL)
(5 mL of brine was added to support the separation). The organic
phase was separated and washed with water (200 mL). This water
phase was extracted with diethyl ether (100 mL). The combined
organic fractions were washed with brine (100 mL), dried over
magnesium sulfate, filtered through a 2 cm layer of silica gel
(silica gel layer was washed with 50 mL of ethyl acetate), and then
concentrated under reduced pressure. The crude product was
subjected to silica gel chromatography (gradient from 0-15% ethyl
acetate in iso-hexanes) to afford the title compound (1.22 g, 67%)
as a colorless oil. R.sub.f=0.48, 30% ethyl acetate in iso-hexanes,
phosphomolybdic acid in ethanol.
##STR00314##
59b (1.01 g, 5 mmol) and copper powder (1.6 g, 25 mmol) were
suspended in tetrahydrofuran (100 mL). Acetic acid (15 mL) was
added and the reaction mixture was heated to 60.degree. C. for 3 h.
After this time, the reaction mixture was poured onto water (copper
powder was filtered off with the use of the filtration paper) and
extracted with diethyl ether (3.times.50 mL). The combined organic
extracts were washed with saturated solution of ammonium chloride
(3.times.50 mL), with saturated solution of sodium bicarbonate
(2.times.50 mL) and with water (50 mL). This water phase was
extracted with diethyl ether (50 mL). Combined organic extracts
were washed with brine (50 mL) and dried over magnesium sulfate,
filtered through a 2 cm layer of silica gel (silica gel layer was
washed with 50 mL of ethyl acetate), and concentrated under reduced
pressure. After drying under high vacuum for one day, the title
compound was isolated (0.84 g, quantitative yield) as a colorless
oil. R.sub.f=0.37, 30% ethyl acetate in iso-hexanes,
phosphomolybdic acid in ethanol.
##STR00315##
Into an oven dried, argon purged flask were placed 59c (0.84 g, 5
mmol) and dichloromethane (2 mL). This mixture was cooled to
0.degree. C. Pinacolborane (0.96 g, 7.5 mmol) was then added
dropwise via syringe. After the mixture was stirred for 1 min., it
was transferred by a syringe into another oven dried, argon purged
flask, immersed in an ice bath and protected from light, containing
zirconocene dichloride (0.13 g, 0.5 mmol). An additional portion of
dichloromethane (2 mL) was used to complete the quantitative
transfer. After this the mixture was warmed to ambient temperature,
it was stirred in the dark for 72 h to achieve full conversion. The
reaction mixture was diluted with ethyl acetate (50 mL) and
carefully quenched with water (1 ml). Water (50 mL) was added and
the organic and aqueous phases were separated. The aqueous phase
was extracted with ethyl acetate (3.times.40 mL). The combined
organic phases were dried over magnesium sulfate, filtered and
evaporated to afford the crude residue which was dissolved in
iso-hexanes (50 mL) and extracted with 5-10% aqueous methanol
(3.times.50 mL) and with brine (50 mL). The hexane phase was dried
over magnesium sulfate. The title compound was isolated as the
white crystalline compound after evaporation (1.42 g, 93%).
R.sub.f=0.38, 30% ethyl acetate in iso-hexanes, iodine vapor.
##STR00316##
Into an oven dried, argon purged flask were placed
(R)-1-(3-chloro-isoquinolin-6-yl)-ethanol (0.83 g, 4 mmol), 59d
(1.40 g, 4.7 mmol), PdCl.sub.2(PCy.sub.2(p-NMe.sub.2Ph)).sub.2
(bis[(dicyclohexyl)(4-dimethylaminophenyl)phosphine]palladium(II)
chloride) (173 mg, 0.21 mmol) and potassium phosphate tribasic
(2.64 g, 12.4 mmol). The flask was sealed with a septum cap and was
re-purged with argon. N,N-Dimethylformamide (10 mL) was added and,
and the resulting reaction mixture was vigorously stirred in an oil
bath pre-heated to 80.degree. C. After 2 h, the reaction was cooled
to ambient temperature and was diluted with ethyl acetate (100 mL)
and water (100 mL). The phases were separated, and the aqueous
phase was extracted with ethyl acetate (2.times.50 mL). The
combined organic phases were dried over magnesium sulfate,
filtered, and concentrated to afford a crude residue that was
purified by silica gel chromatography (gradient from 0-40% ethyl
acetate and methanol (4/1) in iso-hexanes) to afford the title
compound (1.06 g, 78%) as a colorless oil after evaporation.
R.sub.f=0.48, iso-hexanes/ethyl acetate/methanol (6/4/1).
##STR00317##
To a solution of 59e (1.02 g, 3 mmol) in tetrahydrofuran (8 mL) was
added methanol (4 mL), water (4 mL) and lithium hydroxide hydrate
(0.15 g, 6.3 mmol). The resulting mixture was stirred at RT for 10
h and quenched with 1 M hydrochloric acid (6.5 mL, 6.5 mmol). The
resulting solution was concentrated to a crude residue which was
co-evaporated twice with tetrahydrofuran (20 mL), twice with
anhydrous acetonitrile (20 mL) and twice with anhydrous toluene (20
mL). The resulting white solid was dried under high vacuum
overnight and used without further purification (1.24 g,
quantitative yield).
##STR00318##
A solution of 1e (0.53 g, 1 mmol) in dichloromethane (10 mL) was
cooled in an ice water bath. Trimethylsilyl
trifluoromethanesulfonate (0.69 g, 1.80 mmol) was added dropwise at
0.degree. C. under argon, and the resulting solution was stirred at
RT for 30 min. The reaction mixture was evaporated to dryness and
the resulting crude residue was dissolved in anhydrous acetonitrile
(12 mL) under argon. The reaction mixture was stirred at 0.degree.
C., 59f (371 mg, 0.9 mmol) and N,N-diisopropylethylamine (517 mg, 4
mmol) were added followed by
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (570 mg, 1.5 mmol). The reaction
mixture was stirred at RT for 48 h. The solvent was evaporated, the
residue was dissolved in ethyl acetate (50 mL) and the solution was
washed with 20% water solution of citric acid (2.times.50 mL),
water (50 mL) and brine (50 mL), dried over magnesium sulfate,
filtered and evaporated. The residue was purified by silica gel
chromatography (gradient from 0-40% ethyl acetate and methanol
(4/1) in iso-hexanes) to afford the title compound (560 mg, 84%) as
a white solid after evaporation. R.sub.f=0.13, iso-hexanes/ethyl
acetate/methanol (6/4/1).
##STR00319##
To a solution of 59g (371 mg, 0.5 mmol) in tetrahydrofuran (8 mL)
was added methanol (4 mL), water (4 mL) and lithium hydroxide
hydrate (36 mg, 1.5 mmol). The mixture was stirred for 2 h at
ambient temperature and was quenched with aqueous 1 M hydrochloric
acid (1.6 mL, 1.6 mmol). The resulting solution was concentrated to
a crude residue which was co-evaporated twice with tetrahydrofuran
(20 mL), twice with anhydrous acetonitrile (20 mL) and twice with
anhydrous toluene (20 mL). The resulting white solid was dried
under high vacuum overnight and it was used without further
purification (365 mg, quantitative yield). Into oven dried, argon
purged flask were placed 2-methyl-6-nitrobenzoic anhydride (258 mg,
0.75 mmol), 4-dimethylaminopyridine (275 mg, 2.25 mmol) and
anhydrous 1,2-dichloroethane (150 mL). The resulting solution was
heated at 50.degree. C., and the crude seco-acid was added dropwise
via syringe as a solution in dry N,N-dimethylformamide (10 mL) over
12 h. An additional portion of dry N,N-dimethylformamide (2.times.5
mL) was used to complete the quantitative transfer. After stirring
for additional 2 h at 50.degree. C., the reaction mixture was
transferred to a separatory funnel and washed with water (100 mL, 5
mL of brine was added to support the separation). The aqueous phase
was extracted with dichloromethane (50 mL). Combined organic
extracts were washed with brine (50 mL) and dried over magnesium
sulfate, filtered and concentrated under reduced pressure. The
resulting residue was dissolved in ethyl acetate (100 mL) and was
washed with water (100 mL, 5 mL of brine was added to support the
separation). The aqueous phase was extracted with ethyl acetate (50
mL). Combined organic extracts were washed with water (100 mL, 5 mL
of brine was added to support the separation). Resulting aqueous
phase was extracted with ethyl acetate (50 mL). Combined organic
extracts were washed with brine (50 mL) and dried over magnesium
sulfate, filtered and concentrated under reduced pressure. The
residue was purified by silica gel chromatography (gradient from
0-40% ethyl acetate and methanol (4/1) in iso-hexanes) to afford
the title compound (72 mg, 24%) as a white solid after evaporation.
R.sub.f=0.42, 10% methanol in dichloromethane. .sup.1H NMR (400
MHz, CD.sub.3OD): .delta.9.04 (s, 1H), 7.95 (d, J=8.5 Hz, 1H), 7.76
(s, 1H), 7.47 (dd, J=8.5, 1.5 Hz, 1H), 7.31 (s, 1H), 6.43 (m, 2H),
5.93 (q, J=6.6 Hz, 1H), 5.44-5.36 (m, 1H), 4.33-4.23 (m, 2H), 3.84
(dt, J=11.6, 4.1 Hz, 1H), 3.76-3.69 (m, 1H), 3.68-3.61 (m, 2H),
3.48 (m, 1H), 2.65-2.56 (m, 1H), 2.16-2.07 (m, 2H), 2.03-1.93 (m,
1H), 1.91-1.82 (m, 2H), 1.79e (m, 1H), 1.67-1.62 (m, 3H), 1.59 (d,
J=6.6 Hz, 3H), 1.51 (d, J=7.2 Hz, 3H), 0.90 (d, J=6.7 Hz, 3H), 0.85
(d, J=6.7 Hz, 3H). LCMS (m/z) 592.3 [M+H]' Tr=3.12 min.
Example 60
##STR00320##
A solution of Compound 59 (10 mg, 0.017 mmol) in a mixture of ethyl
acetate (4 mL) and tetrahydrofuran (4 mL) containing 10% palladium
on carbon (10 mg) was hydrogenated at RT and at atmospheric
pressure of hydrogen for 3 h. The reaction mixture was filtered
through Celite and the filter pad was washed with tetrahydrofuran
(10 mL). The filtrate was evaporated to afford the title compound
(10 mg, quantitative yield) as a white solid. R.sub.f=0.47, 10%
methanol in dichloromethane. .sup.1H NMR (400 MHz, CD.sub.3OD):
.delta.9.02 (s, 1H), 7.94 (d, J=8.5 Hz, 1H), 7.81 (s, 1H), 7.44 (d,
J=8.5 Hz, 1H), 7.20 (s, 1H), 6.02 (q, J=6.5 Hz, 1H), 5.78-5.71 (m,
1H), 5.24 (m, 1H), 4.33-4.23 (m, 2H), 3.83-3.61 (m, 6H), 3.52 (m,
1H), 2.71-2.66 (m, 1H), 2.43-2.36 (m, 2H), 2.14-2.05 (m, 2H),
2.08-1.91 (m, 1H), 1.90-1.82 (m, 2H), 1.76 (m, 1H), 1.67-1.62 (m,
2H), 1.57 (d, J=6.6 Hz, 3H), 1.54 (d, J=7.2 Hz, 3H), 0.96 (d, J=6.7
Hz, 3H), 0.88 (d, J=6.7 Hz, 3H). LCMS (m/z) 594.3 [M+H]' Tr=2.72
min.
Example 61
##STR00321##
A solution of 1e (1064 mg, 2 mmol) in dichloromethane (30 mL) was
cooled in an ice water bath. Trimethylsilyl
trifluoromethanesulfonate (666 mg, 3 mmol) was added dropwise at
0.degree. C. under argon, and the resulting solution was stirred at
RT for 30 min. The reaction mixture was evaporated to dryness and
the resulting crude residue was dissolved in anhydrous acetonitrile
(25 mL) under argon. The reaction mixture was stirred at 0.degree.
C., hex-5-enoic acid (251 mg, 2.2 mmol) and
N,N-diisopropylethylamine (1034 mg, 8 mmol) were added followed by
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (1065 mg, 2.8 mmol). The reaction
mixture was stirred at RT for 48 h. The solvent was evaporated, the
residue was dissolved in ethyl acetate (200 mL) and the solution
was washed with 20% water solution of citric acid (2.times.150 mL),
water (150 mL) and brine (150 mL), dried over magnesium sulfate,
filtered and evaporated. The residue was purified by silica gel
chromatography (gradient from 0-40% ethyl acetate and methanol
(4/1) in iso-hexanes) to afford the title compound (864 mg, 82%) as
a white solid after evaporation. R.sub.f=0.35, iso-hexanes/ethyl
acetate/methanol (6/4/1).
##STR00322##
To a solution of 61a (830 mg, 1.57 mmol) in tetrahydrofuran (40 mL)
were added water (10 mL) and lithium hydroxide hydrate (57 mg, 2.38
mmol). The mixture was stirred for 2 h at ambient temperature and
then filtered through a 5 cm layer of DOWEX D50.times.8 resin in
H.sup.+ cycle (resin firstly washed with water). Resin was washed
with additional water (50 mL). Filtrates were collected,
concentrated under reduced pressure and co-evaporated twice with
toluene (10 mL). After drying under high vacuum for one day, the
title compound was isolated (590 mg, 95%) as a white solid.
R.sub.f=0.4, 30% methanol in dichloromethane.
##STR00323##
Into an oven dried, argon purged flask were added 61b (238 mg, 0.60
mmol) and (R)-1-(3-vinyl-isoquinolin-6-yl)-ethanol (120 mg, 0.6
mmol). The flask was sealed and the reaction mixture was repurged
twice with argon. Anhydrous dichloromethane (10 mL) was added and
the reaction mixture was repurged twice with argon.
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (148
mg, 0.77 mmol) was added followed by 4-dimethylaminopyridine (67
mg, 0.55 mmol). Reaction mixture was quickly repurged twice with
argon and was stirred at RT for 12 h. The reaction mixture was
diluted with dichloromethane (100 mL) and the solution was washed
with 20% water solution of citric acid (2.times.150 mL), water (150
mL) and brine (150 mL), dried over magnesium sulfate, filtered and
evaporated. The residue was purified by silica gel chromatography
(gradient from 0-40% ethyl acetate and methanol (4/1) in
iso-hexanes) to afford the title compound (307 mg, 89%) as a white
solid after evaporation. R.sub.f=0.29, iso-hexanes/ethyl
acetate/methanol (6/4/1).
##STR00324##
A solution of 61c (209 mg, 0.362 mmol) in toluene (150 mL) was
stirred at RT under argon. Hoveyda-Grubbs 2.sup.nd generation
catalyst (23 mg, 0.036 mmol) was added and the reaction mixture was
heated at reflux under argon for 30 min. Reaction mixture was
cooled to RT and ethyl acetate (50 mL) was added. This solution was
washed twice with aqueous solution of tris(hydroxymethyl)phosphine
(372 mg, 3 mmol in 100 mL of water), with water (2.times.50 mL) and
with brine (50 mL). The organic phase was dried over magnesium
sulfate, filtered and evaporated. The residue was purified by
silica gel chromatography (gradient from 0-40% ethyl acetate and
methanol (4/1) in iso-hexanes) to afford the title compound (125
mg, 63%) as a white solid after evaporation. R.sub.f=0.25,
iso-hexanes/ethyl acetate/methanol (6/4/1). .sup.1H NMR (400 MHz,
CD.sub.3OD): .delta.9.00 (s, 1H), 7.91 (d, J=8.4 Hz, 1H), 7.77 (s,
1H), 7.41 (d, J=8.4 Hz, 1H), 7.23 (s, 1H), 6.64 (m, 1H), 6.29 (d,
J=15.4 Hz, 1H), 5.97 (q, J=5.9 Hz, 1H), 5.53 (q, J=7.0 Hz, 1H),
4.37-4.18 (m, 2H), 3.65 (d, J=9.8 Hz, 1H), 2.65 (t, J=12.3 Hz, 1H),
2.36-2.18 (m, 4H), 1.98 (m, 1H), 1.94-1.73 (m, 3H), 1.71-1.57 (m,
3H), 1.55 (d, J=5.6 Hz, 6H), 0.83 (d, J=6.6 Hz, 3H), 0.78 (d, J=6.6
Hz, 3H). LCMS (m/z) 550.2 [M+H]' Tr=2.55 min.
Example 62
##STR00325##
A solution of Compound 61 (10 mg, 0.018 mmol) in a mixture of ethyl
acetate (4 mL) and tetrahydrofuran (4 mL) containing 10% palladium
on carbon (10 mg) was hydrogenated at RT and at atmospheric
pressure of hydrogen for 3 h. The reaction mixture was filtered
through Celite and the filter pad was washed with tetrahydrofuran
(10 mL). The filtrate was evaporated to afford the title compound
(9 mg, 89%) as a white solid. R.sub.f=0.16, iso-hexanes/ethyl
acetate/methanol (6/4/1). .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.9.02 (s, 1H), 7.93 (d, J=8.7 Hz, 1H), 7.62 (s, 1H), 7.47 (s,
1H), 7.44 (d, J=8.7 Hz, 1H), 6.02-5.93 (m, 1H), 5.39 (m, 1H), 4.03
(m, 1H), 3.69 (m, 1H), 2.84 (m, 2H), 2.27 (s, 1H), 2.10 (m, 1H),
1.97 (m, 1H), 1.85 (m, 2H), 1.76 (m, 2H), 1.62 (m, 4H), 1.54 (d,
J=5.1 Hz, 3H), 1.39 (d, J=6.6 Hz, 3H), 1.19 (m, 4H), 0.78 (d, J=5.0
Hz, 3H), 0.73 (d, J=5.8 Hz, 3H). LCMS (m/z) 552.3 [M+H]' Tr=2.10
min.
Example 63
##STR00326##
A solution of 1e (1064 mg, 2 mmol) in dichloromethane (30 mL) was
cooled in an ice water bath. Trimethylsilyl
trifluoromethanesulfonate (666 mg, 3 mmol) was added dropwise at
0.degree. C. under argon, and the resulting solution was stirred at
RT for 30 min. The reaction mixture was evaporated to dryness and
the resulting crude residue was dissolved in anhydrous acetonitrile
(25 mL) under argon. The reaction mixture was stirred at 0.degree.
C., hept-6-enoic acid (281 mg, 2.2 mmol) and
N,N-diisopropylethylamine (1034 mg, 8 mmol) were added followed by
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (1065 mg, 2.8 mmol). The reaction
mixture was stirred at RT for 48 h. The solvent was evaporated, the
residue was dissolved in ethyl acetate (200 mL) and the solution
was washed twice with 20% water solution of citric acid
(2.times.150 mL), water (150 mL) and brine (150 mL), dried over
magnesium sulfate, filtered and evaporated. The residue was
purified by silica gel chromatography (gradient from 0-40% ethyl
acetate+methanol (4/1) in iso-hexanes) to afford the title compound
(817 mg, 75%) as a white solid after evaporation. R.sub.f=0.37,
iso-hexanes/ethyl acetate/methanol (6/4/1).
##STR00327##
To a solution of 63a (790 mg, 1.46 mmol) in tetrahydrofuran (40 mL)
was added water (10 mL) and lithium hydroxide hydrate (52 mg, 2.19
mmol). The mixture was stirred for 2 h at ambient temperature and
then filtered through a 5 cm layer of DOWEX D50.times.8 resin in
H.sup.+ cycle (resin firstly washed with water). Resin was washed
with additional water (50 mL). The filtrates were collected,
concentrated under reduced pressure and co-evaporated twice with
toluene (10 mL). After drying under high vacuum for one day, the
title compound was isolated (583 mg, 97%) as a white solid.
R.sub.f=0.4, 30% methanol in dichloromethane.
##STR00328##
Into an oven dried, argon purged flask were added 63b (276 mg, 0.67
mmol) and (R)-1-(3-vinyl-isoquinolin-6-yl)-ethanol (134 mg, 0.67
mmol). The flask was sealed and the reaction mixture was repurged
twice with argon. Anhydrous dichloromethane (10 mL) was added and
the reaction mixture was repurged twice with argon.
N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (168
mg, 0.87 mmol) was added followed by 4-dimethylaminopyridine (75
mg, 0.62 mmol). The reaction mixture was quickly repurged twice
with argon and was stirred at RT for 12 h. The reaction mixture was
diluted with dichloromethane (100 mL) and the solution was washed
with 20% water solution of citric acid (2.times.150 mL), water (150
mL) and brine (150 mL), dried over magnesium sulfate, filtered and
evaporated. The residue was purified by silica gel chromatography
(gradient from 0-40% ethyl acetate and methanol (4/1) in
iso-hexanes) to afford the title compound (316 mg, 79%) as a white
solid after evaporation. R.sub.f=0.30, iso-hexanes/ethyl
acetate/methanol (6/4/1).
##STR00329##
A solution of 63c (255 mg, 0.431 mmol) in toluene (200 mL) was
stirred at RT under argon. Hoveyda-Grubbs 2.sup.nd generation
catalyst (27 mg, 0.043 mmol) was added and the reaction mixture was
heated at reflux under argon for 30 min. Reaction mixture was
cooled to RT and ethyl acetate (50 mL) was added. This solution was
washed twice with aqueous solution of tris(hydroxymethyl)phosphine
(372 mg, 3 mmol in 100 mL of water), twice with water (50 mL) and
with brine (50 mL). The organic phase was dried over magnesium
sulfate, filtered and evaporated. The residue was purified by
silica gel chromatography (gradient from 0-40% ethyl acetate and
methanol (4/1) in iso-hexanes) to afford the title compound (76 mg,
31%) as a white solid after evaporation. R.sub.f=0.20,
iso-hexanes/ethyl acetate/methanol (6/4/1). .sup.1H NMR (400 MHz,
CD.sub.3OD): .delta.9.01 (s, 1H), 7.93 (d, J=8.5 Hz, 1H), 7.69 (s,
1H), 7.57 (s, 1H), 7.43 (d, J=8.5 Hz, 1H), 6.55 (m, 1H), 5.95 (m,
1H), 5.43 (m, 1H), 5.23 (m, 3H), 4.32 (m, 1H), 4.20 (m, 1H), 3.67
(m, 1H), 2.69 (m, 1H), 2.30 (m, 2H), 2.13 (m, 2H), 1.96 (m, 3H),
1.83 (m, 1H), 1.64 (m, 3H), 1.56 (d, J=6.7 Hz, 3H), 1.44 (d, J=7.4
Hz, 3H), 0.92-0.72 (m, 6H). LCMS (m/z) 564.4 [M+H], Tr=2.60
min.
Example 64
##STR00330##
A solution of Compound 63 (15 mg, 0.027 mmol) in a mixture of ethyl
acetate (4 mL) and tetrahydrofuran (4 mL) containing 10% palladium
on carbon (10 mg) was hydrogenated at RT and at atmospheric
pressure of hydrogen for 3 h. The reaction mixture was filtered
through Celite and the filter pad was washed with tetrahydrofuran
(10 mL). The filtrate was evaporated to afford the title compound
(13 mg, 87%) as a white solid. R.sub.f=0.11, iso-hexanes/ethyl
acetate/methanol (6/4/1). .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.9.02 (s, 1H), 7.95 (d, J=8.7 Hz, 1H), 7.68 (s, 1H), 7.50 (s,
1H), 7.45 (d, J=8.7 Hz, 1H), 5.93 (q, J=6.3 Hz, 1H), 5.20 (m, 1H),
4.21 (m, 1H), 4.07 (d, J=8.0 Hz, 1H), 3.66 (m, 1H), 2.83 (m, 4H),
2.25 (m, 1H), 2.07 (m, 1H), 1.94 (m, 2H), 1.80e (m, 1H), 1.63 (m,
3H), 1.55 (d, J=6.6 Hz, 3H), 1.51-1.42 (m, 2H), 1.37 (d, J=7.0 Hz,
3H), 1.25 (m, 4H), 0.83 (d, J=6.7 Hz, 6H). LCMS (m/z) 566.3 [M+H],
Tr=2.27 min.
Example 65
##STR00331##
To a solution of (R)-1-(3-chloro-isoquinolin-6-yl)-ethanol (250 mg,
1.21 mmol), and potassium phosphate tribasic (770 mg, 3.63 mmol) in
cyclopentyl methy ether (4.5 mL) and water (1.5 mL) preheated to
90.degree. C. under an argon atmosphere were added
3-(2-methoxy-2-oxoethyl)phenylboronic acid, pinacol ester
(Combi-Blocks, 387 mg, 1.33 mmol) and
(A-.sup.caPhos).sub.2PdCl.sub.2 (49 mg, 60 .mu.mol). After 17 h,
the reaction was allowed to cool to 23.degree. C., and was
partitioned between dichloromethane (50 mL) and saturated aqueous
sodium bicarbonate solution (50 mL). The phases were split and the
aqueous layer was extracted with dichloromethane (50 mL). The
combined organic layers were dried over anhydrous sodium sulfate,
and were concentrated under reduced pressure. The crude residue was
purified by silica gel chromatography (24 g Combiflash HP Gold
Column, 0-100% ethyl acetate/iso-hexanes gradient) to afford the
title compound (321 mg, 83%) as a faint yellow oil.
##STR00332##
To a solution of 65a (320 mg, 1.00 mmol) in tetrahydrofuran (3 mL)
and water (2 mL) was added lithium hydroxide (26 mg, 1.1 mmol) at
23.degree. C. under an argon atmosphere. After 3 h, the reaction
mixture was concentrated under reduced pressure and the crude
residue was purified by silica gel chromatography (24 g Combiflash
HP Gold Column, 0-20% methanol/dichloromethane gradient) to afford
the title compound (64.2 mg, 21%) as a colorless oil. R.sub.f=0.5
(20% methanol in dichloromethane) I.sub.2/silica stain.
##STR00333##
To a solution of 1e (104 mg, 0.21 mmol) in dichloromethane (1.05
mL) was added trimethylsilyl trifluoromethanesulfonate (70 mg, 1.07
mmol) at 0.degree. C. under an argon atmosphere. After 1 h, the
reaction mixture was concentrated under reduced pressure. The
resulting residue was diluted with acetonitrile (1.05 mL) and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (79.8 mg, 0.210 mmol),
N,N-diisopropylethylamine (140 .mu.L, 0.840 mmol), and 65b (64 mg,
0.21 mmol) were sequentially added at 23.degree. C. under an argon
atmosphere. After 18 h, the reaction mixture was concentrated under
reduced pressure, and the crude residue was purified by silica gel
chromatography to afford the title compound (140 mg, 93%) as a
faint yellow oil.
##STR00334##
To a solution of 65c (140 mg, 0.195 mmol) in tetrahydrofuran (0.9
mL) and water (0.3 mL) was added lithium hydroxide hydrate (4.6 mg,
0.195 mmol) at 23.degree. C. under an argon atmosphere. After 3 h,
the reaction mixture was concentrated under reduced pressure to
afford the title compound (131 mg, 99%) as a white solid lithium
carboxylate salt.
##STR00335##
To a solution of benzotriazol-1-yl-oxytripyrrolidinophosphonium
hexafluorophosphate (35.3 mg, 68 .mu.mol) and
4-dimethylaminopyridine (62.3 mg, 510 .mu.mol) in dichloromethane
(5.7 mL) was added 65d (10 mg, 17 .mu.mol) at 23.degree. C. under
an argon atmosphere. After 16 h, the reaction mixture was
concentrated under reduced pressure and the crude residue was
purified by preparative HPLC (Gemini 5u C18 110 .ANG. column,
5-100% acetonitrile/water, 0.1% trifluoroacetic acid modifier) to
afford the title compound (6.0 mg, 52%) as a white solid
trifluoroacetic acid salt. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.9.70 (s, 1H), 8.58 (s, 1H), 8.48 (d, J=8.6 Hz, 1H), 8.32 (s,
1H), 7.97-7.87 (m, 2H), 7.79 (d, J=7.9 Hz, 1H), 7.62 (app t, J=7.7
Hz, 1H), 7.53 (d, J=7.8 Hz, 1H), 6.21 (q, J=6.6 Hz, 1H), 5.84-5.71
(m, 1H), 4.45-4.29 (m, 2H), 3.96 (d, J=15.5 Hz, 1H), 3.75 (dd,
J=11.1, 2.7 Hz, 1H), 3.64 (d, J=15.5 Hz, 1H), 2.74 (td, J=12.8, 3.1
Hz, 1H), 2.13-1.96 (m, 2H), 1.91 (brd, J=13.0 Hz, 1H), 1.81-1.65
(m, 2H), 1.67 (d, J=6.6 Hz, 3H), 1.62 (d, J=7.1 Hz, 3H), 1.01 (d,
J=6.8 Hz, 3H), 0.94 (d, J=6.8 Hz, 3H). HPLC Tr=3.040 min. LCMS
(m/z) 572.3 [M+H], Tr=2.07 min.
Example 66
##STR00336##
To a solution of 6-chloropyridin-3-amine (5.00 g, 38.8 mmol) in
dioxane (194 mL) was added di-tert-butyl dicarbonate (10.2 g, 46.7
mmol) at 23.degree. C. under an argon atmosphere, and the resulting
mixture was heated to 100.degree. C. After 17 h, the reaction
mixture was allowed to cool to 23.degree. C., and was diluted with
water (500 mL). The resulting mixture was extracted with ethyl
acetate (2.times.500 mL), and the combined organic extracts were
dried over anhydrous sodium sulfate and were concentrated under
reduced pressure. The crude residue was purified by silica gel
chromatography (120 g Combiflash HP Gold Column, 0-100% ethyl
acetate/iso-hexanes gradient) to afford the title compound (7.69 g,
87%) as a colorless oil.
##STR00337##
To a solution of 66a (2.00 g, 8.80 mmol) and
tetramethylethylenediamine (2.70 mL, 18.0 mmol) in diethyl ether
(44 mL) was added n-butyllithium (2.5 M in hexanes, 7.2 mL, 18.0
mmol) at -78.degree. C. under an argon atmosphere. After 10 min,
the resulting mixture was allowed to warm to -15.degree. C. over a
50 min period. The reaction mixture was cooled to -78.degree. C.,
and N,N-dimethylformamide (1.9 g, 26 mmol) was added via syringe.
After 30 min, the reaction mixture was quenched with saturated
aqueous ammonium chloride solution (20 mL) and was allowed to warm
to 23.degree. C. The resulting mixture was extracted with ethyl
acetate (2.times.20 mL), and the combined organic layers were dried
over anhydrous sodium sulfate and were concentrated under reduced
pressure. The crude residue was purified by silica gel
chromatography (120 g Combiflash HP Gold Column, 0-100% ethyl
acetate/iso-hexanes gradient) to afford the title compound (849 mg,
38%) as colorless oil.
##STR00338##
To a solution of 66b (258 mg, 1.00 mmol) and
(E)-4-(dimethylamino)but-3-en-2-one (452 mg, 4.00 mmol) in
1,4-dioxane (10 mL) was added p-toluenesulfonic acid monohydrate
(761 mg, 4 mmol) at 23.degree. C. under an argon atmosphere and the
resulting mixture was heated to 80.degree. C. After 2 h, the
reaction mixture was allowed to cool to 23.degree. C. and was
partitioned between saturated aqueous sodium bicarbonate solution
(200 mL) and ethyl acetate (200 mL). The layers were split and the
aqueous layer was extracted with ethyl acetate (200 mL). The
combined organic layers were dried over anhydrous sodium sulfate
and were concentrated under reduced pressure. The crude residue was
purified by silica gel chromatography to afford the title compound
(107 mg, 51%) as an off-white solid.
##STR00339##
Dichloro (p-cymene) ruthenium(II) dimer (5 mg, 8 .mu.mol) and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (7 mg, 19
.mu.mol) were suspended in degassed water (6 mL) and the mixture
was degassed with argon for 15 min. The mixture was stirred at
70.degree. C. under argon for 90 min. The resulting yellow solution
was cooled to RT. 66c (329 mg, 1.56 mmol), sodium formate (543 mg,
7.98 mmol) and degassed tetrahydrofuran (1 mL) were added and the
reaction mixture was degassed for 10 min. The reaction mixture was
vigorously stirred at 40.degree. C. for 2.5 h. The reaction mixture
was cooled to RT and was extracted with ethyl acetate (20 mL). The
organic layer was separated, washed with water (20 mL), brine (20
mL), dried over anhydrous magnesium sulfate, and concentrated under
reduced pressure. The residue was purified by silica gel
chromatography to afford the title compound (180 mg, 54%) as a
solid.
##STR00340##
To a suspension of potassium phosphate tribasic (550 mg, 2.59 mmol)
in cyclopentyl methyl ether (4.5 mL) and water (3 mL) was added 66d
(180 mg, 0.89 mmol) and heated to 90.degree. C. At this
temperature, (A-.sup.caPhos).sub.2PdCl.sub.2(35 mg, 43 .mu.mol) was
added and stirred for 2 min. A solution of
(E)-2,2-dimethyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-but-3-e-
noic acid methyl ester (286 mg, 1.12 mmol) in cyclopentyl methyl
ether (4.5 mL) was added dropwise and stirred for 19 h at
90.degree. C. The reaction mixture was cooled to RT, diluted with
ethyl acetate (40 mL) and water (20 mL). The layers were separated
and the aqueous layer was extracted with ethyl acetate (2.times.20
mL). The combined organic layers were dried over anhydrous
magnesium sulfate, concentrated and the resulting crude residue was
purified via silica gel chromatography (24 g SiO.sub.2 Isco Rf Gold
Column, 0-100% ethyl acetate/iso-hexanes gradient) to afford the
title compound (95 mg, 37%) as a pale brown solid.
##STR00341##
To a solution of 66e (95 mg, 0.32 mmol) in tetrahydrofuran (1.8
mL), methanol (0.6 mL) and water (0.6 mL) was added lithium
hydroxide hydrate (15 mg, 0.63 mmol) at 23.degree. C. After 16 h,
the resulting mixture was concentrated under reduced pressure and
residual solvents were removed azeotropically by addition of
toluene (5 mL) followed by concentration under reduced pressure
(2.times.) to afford the title compound. This was used in the
subsequent amide coupling without further purification.
##STR00342##
To a solution of 1e (212 mg, 0.40 mmol) in dichloromethane (3 mL)
was slowly added trimethylsilyl trifluoromethanesulfonate (106
.mu.L, 0.60 mmol) at 0.degree. C. under an argon atmosphere. After
1 h, the resulting mixture was concentrated under reduced pressure
and was used in the subsequent amide coupling without further
purification. To a solution of 66f (90.5 mg, 0.32 mmol) in
acetonitrile (3 mL),
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (180 mg, 0.47 mmol),
N,N-diisopropylethylamine (330 .mu.L, 1.90 mmol) and
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (232 mg, 0.4
mmol) in acetonitrile (2 mL) were added sequentially at 23.degree.
C. After 16 h, the resulting mixture was concentrated under reduced
pressure and the residue was purified directly by silica gel
chromatography to afford the title compound (201 mg, 91%) as a
solid.
##STR00343##
To a solution of 66g (201 mg, 0.29 mmol) in tetrahydrofuran (2 mL),
methanol (0.4 mL) and water (0.4 mL) was added lithium hydroxide
hydrate (14 mg, 0.57 mmol) at 23.degree. C. After 40 min, lithium
hydroxide hydrate (14 mg, 0.57 mmol) was added at 23.degree. C.
After 1 h, the resulting mixture was concentrated under reduced
pressure and residual solvents were removed azeotropically by
addition of toluene (2 mL) followed by concentration under reduced
pressure (3.times.) to afford the title compound. This was used in
the subsequent macrolactonization without further purification.
##STR00344##
To 2-methyl-6-nitrobenzoic anhydride (43 mg, 0.12 mmol) and
4-dimethylaminopyridine (48 mh, 0.39 mmol) was added
1,2-dichloroethane (20 mL) under nitrogen atmosphere and heated to
50.degree. C. At this temperature, 66h (28 mg, 0.049 mmol) in
N,N-dimethylformamide (1 mL) was added dropwise via syringe pump
over 6 h. An additional wash of N,N-dimethylformamide (0.5 mL) was
then added in the same manner over 15 min. After stirring an
additional 1.25 h, reaction mixture was cool to RT. It was diluted
with ethyl acetate (20 mL) and washed with water (2.times.10 mL).
The organic layer was dried over anhydrous magnesium sulfate and
filtered. The solvent was removed under reduced pressure and the
residue was purified by preparative HPLC (Gemini 5u C18 110 .ANG.
column, 5-100% acetonitrile/water, 0.1% trifluoroacetic acid
modifier) to afford the title compound as a white powder
trifluoroacetic acid salt. It was washed with saturated solution of
sodium bicarbonate to remove acid impurities formed from
2-methyl-6-nitrobenzoic anhydride to afford the title compound (1.5
mg, 5%) as a white solid. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.9.19 (s, 1H), 8.86 (d, J=2.1 Hz, 1H), 8.67 (d, J=7.5 Hz,
1H), 8.21 (s, 1H), 7.59 (s, 1H), 6.89 (d, J=9.5 Hz, 1H), 6.58 (d,
J=16.1 Hz, 1H), 6.46 (d, J=16.1 Hz, 1H), 6.04 (q, J=6.4 Hz, 1H),
5.52-5.41 (m, 1H), 4.26 (d, J=14.4 Hz, 1H), 4.18 (app t, J=9.1 Hz,
1H), 3.65 (dd, J=11.4, 2.8 Hz, 1H), 2.58 (td, J=12.9, 3.2 Hz, 1H),
1.91-1.75 (m, 3H), 1.68-1.53 (m, 2H), 1.63 (d, J=6.7 Hz, 3H), 1.51
(d, J=7.3 Hz, 3H), 1.43 (s, 3H), 1.26 (s, 3H), 0.87 (d, J=6.7 Hz,
3H), 0.82 (d, J=6.7 Hz, 3H).LCMS (m/z) 551.2 [M+H], Tr=2.19
min.
Example 67
##STR00345##
Methyl 2-(3-bromophenyl)-2-methylpropanoate (Pharmabridge,
Doylestown, Pa., USA (190 mg, 0.74 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (206
mg, 0.81 mmol),
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II),
complex with dichloromethane (27 mg, 0.04 mmol) and potassium
acetate (217 mg, 2.22 mmol) were placed in a screw cap vial and
flushed with a vacuum and argon cycle three times. Anhydrous
1,4-dioxane (4 mL) was added under argon and the resulting mixture
was heated to 80.degree. C. for 19 h. The reaction mixture was cool
to RT and was diluted with ethyl acetate (20 mL). Celite (.about.1
g) was added and filtered through a pad of celite. Solvents were
removed under reduced pressure to afford the title compound which
was used directly in the subsequent reaction.
##STR00346##
To a suspension of potassium phosphate tribasic (445 mg, 2.10 mmol)
in cyclopentyl methyl ether (2.5 mL) and water (1.5 mL) was added
(R)-1-(3-chloro-isoquinolin-6-yl)-ethanol (180 mg, 0.89 mmol) and
the reaction mixture was heated to 90.degree. C. At this
temperature, (A-.sup.caPhos).sub.2PdCl.sub.2 (28 mg, 35 .mu.mol)
was added and the reaction mixture was stirred for 2 min. A
solution of 67a (225 mg, 0.74 mmol) in cyclopentyl methyl ether
(2.5 mL) was added dropwise and stirred for 19 h at 90.degree. C.
The reaction mixture was cool to RT, diluted with ethyl acetate (30
mL) and water (15 mL). The layers were separated and the aqueous
layer was extracted with ethyl acetate (2.times.15 mL). The
combined organic layers were dried over anhydrous magnesium
sulfate, concentrated and the resulting crude residue was purified
by silica gel column chromatography to afford the title compound
(267 mg, quantitative) as a pale brown solid.
##STR00347##
To a solution of 67b (244 mg, 0.7 mmol) in tetrahydrofuran (10 mL),
and water (2.5 mL) was added lithium hydroxide hydrate (18.5 mg,
0.77 mmol) at 23.degree. C. After 48 h, lithium hydroxide hydrate
(17 mg, 0.70 mmol) was added to the reaction mixture. After 24 h,
the reaction mixture was concentrated under reduced pressure and
residual solvents were removed azeotropically by addition of
toluene (5 mL) followed by concentration under reduced pressure
(2.times.) to afford the title compound. This was used in the
subsequent amide coupling without further purification.
##STR00348##
To a solution of 1e (397 mg, 0.75 mmol) in dichloromethane (5 mL)
was slowly added trimethylsilyl trifluoromethanesulfonate (199
.mu.L, 1.12 mmol) at 0.degree. C. under an argon atmosphere. After
1 h, the resulting mixture was concentrated under reduced pressure
to give
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester as a triflate
salt. This was used in the subsequent amide coupling without
further purification. To a solution of 67c (234 mg, 0.7 mmol) in
acetonitrile (7 mL),
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (532 mg, 1.4 mmol),
N,N-diisopropylethylamine (730 .mu.L, 4.2 mmol) and
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (435 mg, 0.75
mmol) in acetonitrile (3 mL) were added sequentially at 23.degree.
C. After 16 h, the resulting mixture was concentrated under reduced
pressure and the residue was purified directly by silica gel
chromatography (40 g SiO.sub.2 Isco Rf Gold Column, 0-100% ethyl
acetate/iso-hexanes gradient) to afford the title compound (230 mg,
44%) as a solid.
##STR00349##
To a solution of 67d (94 mg, 0.13 mmol) in tetrahydrofuran (3 mL),
and water (1 mL) was added lithium hydroxide hydrate (3.1 mg, 0.13
mmol) at 23.degree. C. After 48 h, lithium hydroxide hydrate (17
mg, 0.70 mmol) was added to the reaction mixture. After 1 h, the
reaction mixture was concentrated under reduced pressure and
residual solvents were removed azeotropically by addition of
toluene (5 mL) followed by concentration under reduced pressure
(2.times.) to afford the title compound. This was used in the
subsequent macrolactonization without further purification.
##STR00350##
To a suspension of 67e (78 mg, 0.13 mmol) in dichloromethane (42
mL) was added benzotriazol-1-yl-oxytripyrrolidinophosphonium
hexafluorophosphate (520 mg, 0.50 mmol) and 4-dimethylaminopyridine
(462 mg, 4.0 mmol) at 23.degree. C. After 24 h, the solvent was
removed under reduced pressure and the residue was purified by
preparative HPLC (Gemini 5u C18 110 .ANG. column, 5-100%
acetonitrile/water, 0.1% trifluoroacetic acid modifier) to afford
the title compound (3.2 mg, 4%) as a white powder and as a
trifluoroacetic acid salt. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.9.65 (s, 1H), 8.69 (s, 1H), 8.49 (d, J=8.7 Hz, 1H), 8.44 (d,
J=8.6 Hz, 1H), 8.34 (s, 1H), 7.90 (dd, J=8.7, 1.5 Hz, 1H), 7.83 (d,
J=7.7 Hz, 1H), 7.78 (d, J=7.9 Hz, 1H), 7.74 (s, 1H), 7.69 (app t,
J=7.8 Hz, 1H), 6.82 (d, J=8.6 Hz, 1H), 6.21 (q, J=6.5 Hz, 1H),
5.92-5.84 (m, 1H), 4.41-4.30 (m, 2H), 3.75 (dd, J=14.4, 6.0 Hz,
1H), 2.71 (td, J=13.0, 3.0 Hz, 1H), 2.06-1.99 (m, 1H), 1.96 (dd,
J=14.2, 7.1 Hz, 1H), 1.93-1.86 (m, 1H), 1.75 (s, 3H), 1.73-1.59 (m,
2H), 1.68 (d, J=6.6 Hz, 3H), 1.63 (d, J=7.1 Hz, 3H), 1.47 (s, 3H),
0.96 (d, J=6.8 Hz, 3H), 0.84 (d, J=6.7 Hz, 3H). HPLC Tr=5.319 min.
LCMS (m/z) 600.5 [M+H], Tr=2.71 min.
Examples 68 and 69
##STR00351##
To a solution of 6-bromo-3-chloroisoquinoline (Frontier Scientific,
1.594 g, 6.57 mmol) and 2,2-dimethylpropane-1,3-diol (684 mg, 6.57
mmol) in cyclopentyl methyl ether (20 mL) was added cesium
carbonate (2.354 g, 7.23 mmol) at 23.degree. C. The reaction
mixture was heated to 120.degree. C. for 18 h. The reaction mixture
was cool to RT, diluted with ethyl acetate (50 mL) and washed with
water (30 mL), brine (30 mL) and the resulting organic layer was
dried over anhydrous magnesium sulfate and concentrated under
reduced pressure. The residue was purified by silica gel
chromatography (80 g Isco Rf Gold Column, 0-100% ethyl
acetate/iso-hexanes gradient) to afford the title compound (461 mg,
23%) as a white solid.
##STR00352##
To a solution of 68a (461 mg, 1.49 mmol) in acetonitrile (10 mL)
and water (2.5 mL) was added periodic acid (1.698 g, 7.45 mmol) at
23.degree. C. The reaction mixture was cool to 0.degree. C. and
chromium trioxide (30 mg, 0.298 mmol) was added in one portion.
After 2.5 hour, the reaction mixture was diluted with ethyl acetate
(30 mL) and water (30 mL) and the layers were separated. The
aqueous layer was extracted with ethyl acetate (30 mL) and combined
organic layers were washed with brine, dried over anhydrous
magnesium sulfate and concentrated under reduced pressure to afford
the title compound which was used without purification (481
mg).
##STR00353##
To a solution of 68b (481 mg, 1.49 mmol) in acetonitrile (10 mL),
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (1.133 g, 2.98 mmol),
N,N-diisopropylethylamine (1.55 mL, 8.94 mmol) and (S)-methyl
2-amino-3-methylbutanoate hydrochloride (749 mg, 4.47 mmol) were
added sequentially at 23.degree. C. After 16 h, the resulting
mixture was concentrated under reduced pressure and the residue was
purified directly by silica gel chromatography (24 g Isco Rf Gold
Column, 0-100% ethyl acetate/iso-hexanes gradient) to afford the
title compound (412 mg, 63% over 2 steps) as a brown oil.
##STR00354##
To a suspension of 68c (412 mg, 0.95 mmol) and
bis(triphenylphosphine) palladium (II) dichloride in toluene (5 mL)
was added tributyl(1-ethoxyvinyl)tin (962 .mu.L, 2.85 mmol) under
argon at 23.degree. C. The reaction mixture was heated to
50.degree. C. After 18 h, water (1 mL) was added at 50.degree. C.
After 2 h, the reaction mixture was cool to RT, diluted with ethyl
acetate (15 mL) and 1 M solution of potassium fluoride (5 mL) was
added. The resulting mixture was vigorously stirred at 23.degree.
C. After 2 h, the mixture was filtered through a short pad of
Celite and washed with ethyl acetate (10 mL). The filtrate was
washed with water (15 mL), brine (15 mL), dried over anhydrous
magnesium sulfate and concentrated. This residue was dissolved in
1,4-dioxane (8 mL) and 1 M aqueous hydrochloric acid (1 mL) was
added at 23.degree. C. After 5 min, the reaction mixture was
quenched with a saturated solution of sodium bicarbonate (2 mL) and
concentrated to dryness. The residue was taken up in ethyl acetate
(20 mL) and water (20 mL) and layers were separated. The aqueous
layer was extracted with ethyl acetate (2.times.10 mL) and the
combined organic layers were washed with brine (20 mL), dried over
anhydrous magnesium sulfate and concentrated. The residue was
purified by silica gel chromatography to afford the title compound
(236 mg, 62%) as a brown gum.
##STR00355##
Dichloro (p-cymene) ruthenium(II) dimer (2 mg, 3 .mu.mol) and
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (2.6 mg, 7
.mu.mol) were suspended in degassed water (2.5 mL) and the mixture
was degassed with argon for 15 min. The mixture was stirred at
70.degree. C. under nitrogen for 90 min. The resulting yellow
solution was cooled to RT. 68d (236 mg, 0.59 mmol), sodium formate
(200 mg, 2.95 mmol) and degassed tetrahydrofuran (1.25 mL) were
added and the reaction mixture was degassed for 10 min. The
reaction mixture was vigorously stirred at 40.degree. C. for 3.5 h.
The reaction mixture was cooled to RT and was extracted with ethyl
acetate (15 mL). The organic layer was separated, washed with water
(20 mL), brine (20 mL), dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure. The residue was purified by
silica gel chromatography to afford the title compound (196 mg,
83%) as a pale brown solid.
##STR00356##
To a solution of 68e (196 mg, 0.487 mmol) in tetrahydrofuran (3
mL), methanol (1 mL) and water (1 mL) was added lithium hydroxide
hydrate (23 mg, 0.97 mmol) at 23.degree. C. After 16 h, the
reaction mixture was concentrated under reduced pressure. The
residue was taken up in ethyl acetate (10 mL) and water (10 mL) and
acidified with 1 M aqueous hydrochloric acid solution to pH
.about.2. The resulting layers were separated. The aqueous layer
was extracted with ethyl acetate (2.times.10 mL) and the combined
organic layers were washed with brine, dried over anhydrous
magnesium sulfate and concentrated under reduced pressure to afford
the title compound which was used without purification.
##STR00357##
To a solution of
(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-c-
arboxylic acid 2,2,2-trichloro-ethyl ester (270 mg, 0.62 mmol) in
dichloromethane (5 mL) was slowly added trimethylsilyl
trifluoromethanesulfonate (165 .mu.L, 0.93 mmol) at 0.degree. C.
under an argon atmosphere. After 1 hour, the reaction mixture was
concentrated under reduced pressure to afford the triflate salt of
(S/R)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic
acid 2,2,2-trichloro-ethyl ester as yellow oil which was used it
without purification. To a solution of 68f (189 mg, 0.49 mmol) in
acetonitrile (7 mL),
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (278 mg, 0.73 mmol),
N,N-diisopropylethylamine (730 .mu.L, 4.2 mmol) and
(S/R)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic
acid 2,2,2-trichloro-ethyl ester (435 mg, 0.62 mmol) in
acetonitrile (3 mL) were added sequentially at 23.degree. C. After
16 h, the resulting mixture was concentrated under reduced pressure
and the residue was purified directly by silica gel chromatography
(40 g SiO.sub.2 Isco Rf Gold Column, 0-100% ethyl
acetate/iso-hexanes gradient) to afford an inseparable
diastereomeric mixture of the title compounds (43 mg, 13%) as a
colorless residue.
##STR00358##
To a solution of a mixture of 68g and 68h (43 mg, 61 .mu.mol) in
tetrahydrofuran (4 mL) and water (2 mL) was added lithium hydroxide
hydrate (3.2 mg, 0.13 mmol) at 23.degree. C. After 1 h, the
reaction mixture was concentrated under reduced pressure and
residual solvents were removed azeotropically by addition of
toluene (5 mL) followed by concentration under reduced pressure
(2.times.) to afford the title compounds as a mixture. This was
used in the subsequent macrolactonization without further
purification. LCMS (m/z) 572.2 [M+H], Tr=2.21 min.
##STR00359##
To a suspension of 68i and 68j (34.8 mg, 61 .mu.mol) in
dichloromethane (20 mL) was added
benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate
(127 mg, 0.24 mmol) and 4-dimethylaminopyridine (224 mg, 1.83 mmol)
at 23.degree. C. After 24 h, the solvent was removed under reduced
pressure and the residue was purified by preparative HPLC (Gemini
5u C18 110 .ANG. column, 5-100% acetonitrile/water, 0.1%
trifluoroacetic acid modifier) to afford both compounds (12 mg,
35%) as a ratio mixture of diastereomers. The diastereomers were
separated using Chiral preparative HPLC to afford first eluting
(Tr=3.73 min) Compound 68 (2.17 mg, 6%) and the second eluting
(Tr=5.713 min)
Compound 69 (1.85 mg, 5%). Compound 68: .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta.8.89 (s, 1H), 7.94 (d, J=8.6 Hz, 1H), 7.86 (s,
1H), 7.37 (dd, J=8.6, 1.5 Hz, 1H), 7.28 (d, J=9.2 Hz, 1H), 6.93 (s,
1H), 6.09 (q, J=6.5 Hz, 1H), 5.54 (q, J=7.2 Hz, 1H), 4.36-4.26 (m,
3H), 4.21 (d, J=9.7 Hz, 1H), 3.69 (dd, J=11.2, 2.6 Hz, 1H), 2.77
(td, J=12.9, 3.0 Hz, 1H), 2.00-1.84 (m, 2H), 1.83-1.77 (m, 1H),
1.76-1.62 (m, 2H), 1.67 (d, J=6.7 Hz, 3H), 1.47 (d, J=7.3 Hz, 3H),
1.38 (s, 3H), 1.27 (s, 3H), 0.99 (d, J=6.7 Hz, 3H), 0.95 (d, J=6.7
Hz, 3H). HPLC Tr=5.866 min. LCMS (m/z) 554.2 [M+H], Tr=2.44
min.
Compound 69: .sup.1H NMR (400 MHz, CD3OD) .delta. 8.78 (s, 1H),
7.83 (d, J=8.6 Hz, 1H), 7.74 (s, 1H), 7.24 (dd, J=8.6, 1.6 Hz, 1H),
7.03 (s, 1H), 5.93 (q, J=6.7 Hz, 1H), 5.35 (q, J=6.8 Hz, 1H), 4.31
(d, J=9.8 Hz, 1H), 4.18 (d, J=13.3 Hz, 1H), 4.02-3.97 (m, 2H), (d,
J=10.0 Hz, 1H), 3.65 (dd, J=10.8, 2.8 Hz, 1H), 2.71 (td, J=12.7,
2.8 Hz, 1H), 1.98-1.89 (m, 1H), 1.87-1.77 (m, 2H), 1.73-1.64 (m,
1H), 1.57 (d, J=6.7 Hz, 3H), 1.49 (d, J=7.0 Hz, 3H), 1.40 (s, 3H),
1.13 (s, 3H), 0.89 (d, J=6.7 Hz, 3H), 0.85 (d, J=6.5 Hz, 3H) HPLC
Tr=5.951 min. LCMS (m/z) 555.2 [M+H], Tr=2.49 min.
Example 70
##STR00360##
To 2,7-dibromonaphthalene (1 g, 3.50 mmol) in anhydrous
tetrahydrofuran (18 mL), at -78.degree. C. and under an atmosphere
of nitrogen, was added a solution of n-butyllithium (2.5 M in
hexanes, 1.5 mL, 3.67 mmol) dropwise. The reaction was stirred at
-78.degree. C. for 20 min after which N-methoxy-N-methylacetamide
(409 .mu.L, 3.85 mmol) was added. After 15 min, the reaction was
warmed to RT and stirred for 30 min. The reaction was quenched with
2 M hydrochloric acid and extracted twice with dichloromethane. The
combined organic layers were dried through a hydrophobic frit and
concentrated in vacuo. The product was purified by silica flash
chromatography (iso-hexanes/ethyl acetate, 7/1) to afford the title
compound (650 mg, 75%) as a colorless solid.
##STR00361##
To dichloro(p-cymene)ruthenium(II)dimer(8 mg, 0.013 mmol) in water
(5 mL) at RT was added
(1R,2R)-(-)-N-p-tosyl-1,2-diphenylethylenediamine (11.5 mg, 0.031
mmol). The system was degassed for 15 min and then heated to
70.degree. C. for 1.5 h. The reaction was cooled and was added a
solution of 70a (650 mg, 2.61 mmol) in degassed anhydrous
tetrahydrofuran (2 mL) followed by sodium formate (874 mg, 13.1
mmol). The reaction was heated at 40.degree. C. for 3 h, cooled to
RT and extracted twice with dichloromethane. The combined organic
layers were dried through a hydrophobic frit and concentrated in
vacuo. The product was purified by silica gel chromatography
(iso-hexanes/ethyl acetate, 4/1) to afford the title compound (450
mg, 69%) as a colorless solid.
##STR00362##
70b (42 mg, 0.17 mmol) was dissolved in acetonitrile (2 mL) in a
microwave vial, to the mixture was added 3-butenoic acid (35 mg,
0.41 mmol), palladium (II) acetate (4 mg, 0.017 mmol),
tri-(o-tolyl)phosphine (10 mg, 0.034 mmol) and triethylamine (0.12
mL). The vial was heated at 120.degree. in the microwave reactor
for 15 mins. The reaction mixture was then filtered, the solvent
was evaporated and purified with combi-flash column chromatography
(Eluent methanol/dichloromethane 1:3) to afford the title compound
(35 mg, 81%) as a yellow solid.
##STR00363##
70c (40 mg, 0.08 mmol) was dissolved in N,N-dimethylformamide (2
mL), to the solution was added
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (36 mg, 0.1 mmol), the reaction
mixture was stirred at RT for 10 mins. Then
(S)-1-[(S)-2-((S)-2-amino-3-methylbutyrylamino)-propionyl]-hexahydro-pyri-
dazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester (0.18 mmol) in
N,N-dimethylformamide (1 mL) was added to above reaction mixture
followed by triethylamine (32 mg, 0.32 mmol). The reaction mixture
was stirred at RT for 1 h, and then it was diluted with ethyl
acetate (20 mL) and washed with brine. The aqueous layer was back
extracted with ethyl acetate and the combined organic solvent was
evaporated and purified with combi-flash column chromatography
(Eluent methanol/dichloromethane 1:10) to afford the title compound
(68 mg, 65%) as a yellow solid.
##STR00364##
70d (55 mg, 0.08 mmol) was dissolved in the mixture of
tetrahydrofuran (2 mL), methanol (1 mL) and water (1 mL). To the
solution was added lithium hydroxide hydrate (4 mg, 0.16 mmol). The
reaction mixture was stirred at RT for 1 h. Dichloromethane (10 mL)
and water (10 mL) were added to the reaction mixture. 1 N
Hydrochloric acid was added to the aqueous layer until pH reached
2, the acidic aqueous layer was extracted with dichloromethane
(2.times.10 mL). The organic solvent was then evaporated to afford
the title compound (37 mg, 84%) as a white solid.
##STR00365##
2-Methyl-6-nitrobenzoic anhydride (19 mg, 0.056 mmol) and
4-dimethylaminopyridine (23 mg, 0.185 mmol) were dissolved in
1,2-dichloroethane (18 mL) and the solution was heated at
50.degree. C. To the above solution was added 70e (20 mg, 0.037
mmol) in N,N-dimethylformamide (1 mL) via syringe pump in 10 h. The
reaction mixture was stirred at 50.degree. C. for 2 h after the
completion of addition. The solvent was then evaporated and the
residue was purified by reverse phase preparative HPLC (0-100%
acetonitrile/water) to afford the title compound (4 mg, 22%) as a
white solid. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.7.79-7.72 (m,
3H), 7.52 (dd, J=1.6, 8.8 Hz, 1H), 7.41 (s, 1H), 7.33 (dd, J=8.4,
1.6 Hz, 1H), 6.48 (d, J=16.4 Hz, 1H), 6.31-6.24 (m, 1H), 6.02 (q,
J=7.6 Hz, 1H), 4.64 (d, J=12.4 Hz, 1H), 4.40 (d, J=12.8 Hz, 1H),
4.30 (d, J=10.4 Hz, 1H), 3.78-3.72 (m, 1H), 3.36-3.32 (m, 2H),
2.99-2.93 (m, 1H), 2.77-2.71 (m, 1H), 1.99-1.69 (m, 5H), 1.66 (d,
J=6.8 Hz, 3H), 1.58 (d, J=7.2 Hz, 3H), 0.99 (d, J=6.8 Hz, 6H). LCMS
(m/z) 521.1 [M-H], Tr=3.18 min.
Example 71
##STR00366##
Compound 70 (5 mg, 0.01 mmol) was dissolved in ethanol (5 mL) under
argon, to the solution was added Pd (10% on activated carbon, 3
mg). The reaction flask was then purged and then charged with
H.sub.2 using a balloon. The reaction was filtered through Celite
after 2 h, the filtrate was evaporated under reduced pressure and
purified by reverse phase preparative HPLC (0-100%
acetonitrile/water) to afford the title compound (3.2 mg, 64%) as a
white powder. .sup.1H-NMR (400 MHz, CD.sub.3OD) .delta.8.47 (d,
J=6.0 Hz, 1H), 7.97 (d, J=8.8 Hz, 1H), 7.69-7.62 (m, 3H), 7.31 (s,
1H), 7.24-7.19 (m, 2H), 5.97 (dd, J=12.8, 6.0 Hz, 1H), 5.62 (pent,
J=7.2 Hz, 1H), 4.76-4.16 (m, 1H), 4.10 (app t, J=12.8 Hz, 1H),
4.05-3.69 (m, 1H), 2.85 (br s, 1H), 2.73-2.41 (m, 2H), 2.40-2.37
(m, 1H), 2.10-1.93 (m, 1H), 1.92-1.70 (m, 6H), 1.53 (d, J=6.8 Hz,
3H), 1.49 (d, J=7.6 Hz, 3H), 0.99 (d, J=6.8 Hz, 6H). LCMS (m/z)
523.140 [M+H], Tr=3.16 min.
Example 72
##STR00367##
Into an oven dried, argon purged flask Compound 49 (10 mg, 0.017
mmol), copper(I) iodide (1 mg, 0.005 mmol) and prop-2-yn-1-ol (4
mg, 0.07 mmol) were added. The flask was sealed and repurged with
argon three times. Anhydrous N,N-dimethylformamide (5 mL) was added
and the reaction mixture was repurged with argon three times. This
reaction mixture was stirred at RT for 12 h. After evaporation of
the solvent under reduced pressure, the crude residue was dissolved
in ethyl acetate (10 mL) and filtered through filter aid and the
filter pad was washed with ethyl acetate (10 mL). After
concentration under reduced pressure, the residue was purified by
silica gel chromatography (gradient from 0-40% ethyl acetate and
methanol (4/1) in iso-hexanes) to afford the title compound (10 mg,
93%) as a white solid after evaporation. R.sub.f=0.33, 10% methanol
in dichloromethane. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.9.05
(s, 1H), 7.95 (d, J=8.6 Hz, 1H), 7.94 (s, 1H), 7.82 (s, 1H), 7.50
(s, 1H), 7.46 (d, J=8.6 Hz, 1H), 6.49 (d, J=16.0 Hz, 1H), 6.40 (d,
J=16.0 Hz, 1H), 5.95 (m, 1H), 5.50 (m, 1H), 4.59-4.53 (m, 3H), 4.36
(m, 2H), 4.30 (m, 1H), 3.68 (m, 1H), 2.80 (m, 1H), 2.60 (m, 1H),
2.23 (m, 1H), 2.14 (m, 1H), 1.90-1.58 (m, 4H), 1.57 (d, J=6.6 Hz,
3H), 1.55 (d, J=7.6 Hz, 3H), 1.41 (s, 3H), 1.27 (s, 3H). LCMS (m/z)
633.4 [M+H], Tr=2.66 min.
Example 73
##STR00368##
To 3-Nitromethylene-oxetane (Angew. Chem. Int. Ed. 2006, 45 (46),
7736, 2.5 g, 21.7 mmol) in anhydrous tetrahydrofuran (40 mL) at RT
and under an atmosphere of nitrogen was added copper(I) iodide (413
mg, 2.17 mmol) and chlorotrimethylsilane (3.0 mL, 23.9 mmol). The
resulting yellow solution was stirred for 5 min and cooled to
between -15.degree. C. and -11.degree. C. with a methanol ice bath.
Vinyl magnesium bromide (43.5 mL, 43.5 mmol, 1.0 M in
tetrahydrofuran) was slowly added over 3 h via a syringe pump.
Following the addition the reaction was quenched with a saturated
aqueous solution of ammonium chloride, filtered and extracted with
diethyl ether (3.times.). The combined organics were dried through
a hydrophobic frit and concentrated in vacuo. The residue was
purified by silica gel chromatography using diethyl ether to give
the title compound as a yellow oil (2.0 g, 64%).
##STR00369##
To 73a (600 mg, 4.2 mmol) in anhydrous dimethylsulfoxide (13 mL)
was added acetic acid (2.4 mL, 42 mmol) and sodium nitrite (869 mg,
12.6 mmol) and the mixture heated to 35.degree. C. for 16 h. The
reaction was cooled to RT and diluted with water. The pH was
adjusted to pH 3-4 with 10% aqueous hydrochloric acid and the
product extracted with diethyl ether (3.times.) and diethyl
ether/ethyl acetate (1:1). The combined organics were dried through
a hydrophobic frit and concentrated in vacuo. The residue was
purified by silica gel chromatography using iso-hexanes/ethyl
acetate (2/1 then 1/1) to afford impure acid. This was dissolved in
diethyl ether and extracted with a saturated solution of sodium
carbonate. The aqueous was acidified to pH 2 with concentrated
hydrochloric acid and extracted with ethyl acetate (3.times.). The
organics were dried through a hydrophobic frit and concentrated in
vacuo to give the title compound as a yellow oil (228 mg, 43%).
##STR00370##
To 73b (150 mg, 1.18 mmol) in anhydrous dioxane (1 mL), was added
acetic acid (R)-1-(7-bromo-quinolin-2-yl)-ethyl ester (347 mg, 1.18
mmol) followed by dicyclohexylmethylamine (0.76 mL, 3.54 mmol),
palladium(II) acetate (53 mg, 0.24 mmol) and Tri(o-tolyl) phosphine
(72 mg, 0.24 mmol). The mixture was heated at 100.degree. C. for 1
hour. An additional amount of palladium(II) acetate (26 mg, 0.12
mmol) and Tri(o-tolyl) phosphine (36 mg, 0.12 mmol) was added and
heating continued at 100.degree. C. for a further 45 min. The
reaction was cooled to RT and 2M HCl added until pH 3-4 was
reached. The product was extracted with ethyl acetate (3.times.)
and ethyl acetate/10% methanol (2.times.). The combined organics
were dried through a hydrophobic frit and concentrated in vacuo to
yield the title compound as a brown oil.
##STR00371##
A solution of 1e (3.01 g, 5.66 mmol) in tetrahydrofuran:methanol
(1:1, 60 mL) was stirred at 0.degree. C. Tetra-n-butylammonium
fluoride (1 M in tetrahydrofuran, 11.3 mL, 11.3 mmol) was added and
the reaction mixture was stirred at RT for 22 h. The solvent was
evaporated and the residue was purified by silica gel
chromatography using iso-hexane to iso-hexane/ethyl acetate 1:1
afford the title compound (2.14 g, 91%) as a white foam.
##STR00372##
To 73d (365 mg, 0.88 mmol) in anhydrous dichloromethane (15 mL) at
0.degree. C. and under an atmosphere of nitrogen, was added
trimethylsilyl trifluoromethanesulfonate (239 .mu.L, 1.32 mmol).
The reaction mixture was stirred at 0.degree. C. for 1 hour before
adding N,N-diisopropylethylamine (613 .mu.L, 3.52 mmol) and then
concentrated in vacuo, and co-evaporated with toluene to afford
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester as a white
solid. To 73c (300 mg, 0.88 mmol) in anhydrous acetonitrile (9 mL)
at 0.degree. C. and under an atmosphere of nitrogen was added
N,N-diisopropylethylamine (766 .mu.L, 4.4 mmol) and
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (401 mg, 1.06 mmol). The solution
was stirred at 0.degree. C. for 3 min before adding a solution of
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid 2,2,2-trichloro-ethyl ester in anhydrous
acetonitrile (2 mL). The reaction was warmed to RT stirred for 2 h.
The reaction was quenched with 1M HCl and extracted with ethyl
acetate (3.times.). The combined organic layers were dried through
a hydrophobic frit and concentrated in vacuo. The residue was
purified by silica gel chromatography using ethyl acetate to give
the title compound as a viscous yellow oil (160 mg, 22%, 2
steps).
##STR00373##
To 73e (160 mg, 0.25 mmol) in tetrahydrofuran (10 mL) and water (2
mL) was added lithium hydroxide monohydrate (53 mg, 1.25 mmol) at
0.degree. C. The reaction was stirred at 0.degree. C. for 2 h and
quenched by adding 2M aqueous hydrochloric acid (0.63 mL). The
reaction was concentrated in vacuo, followed by co-evaporation from
toluene/methanol (3.times.) and then toluene (3.times.) and dried
on a high vacuum for 15 min. The resulting residue was dissolved in
anhydrous tetrahydrofuran (83 mL) and at RT was added
N,N-diisopropylethylamine (223 .mu.L, 1.25 mmol),
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (114 mg, 1.3 mmol) and
4-dimethylaminopyridine (3 mg, 0.03 mmol). The reaction was stirred
for 24 h, diluted with ethyl acetate and washed with 1M HCl
(1.times.) and brine (1.times.). The organic layer was dried
through a hydrophobic frit and concentrated in vacuo. The residue
was purified by silica gel chromatography using ethyl
acetate/methanol 1:0 then 20/1 to give a viscous yellow oil (52
mg). This was further purified by preparative thin layer
chromatography (ethyl acetate) to afford the title compound as a
white solid which was triturated with diethyl ether, filtered and
vacuum dried. (22 mg, 16%, 2 steps). .sup.1H NMR (300 MHz,
CDCl.sub.3) 0.96 (d, J=6.5 Hz, 3H), 1.00 (d, J=6.5 Hz, 3H), 1.50
(d, J=7.1 Hz, 3H), 1.69 (d, J=6.9 Hz, 3H), 1.81-2.16 (m, 4H),
2.63-2.74 (m, 1H), 3.26-3.38 (m 1H), 3.70-3.83 (m, 1H), 3.98 (d,
J=12.3 Hz, 1H), 4.28 (t, J=9.8 Hz, 1H), 4.33-4.43 (m, 1H), 4.57 (d,
J=5.8 Hz, 1H), 4.86 (q, J=6.7 Hz, 2H), 5.13 (d, J=5.8 Hz, 1H), 5.73
(t, J=6.9 Hz, 1H), 5.96 (q, J=6.7 Hz, 1H), 6.24 (d, J=16.3 Hz, 1H),
6.61-6.73 (m, 1H), 7.00 (d, J=16.3 Hz, 1H), 7.09-7.20 (m, 1H), 7.40
(d, J=8.5 Hz, 1H), 7.58 (s, 1H), 7.81-7.93 (m, 2H), 8.22 (d, J=8.7
Hz, 1H). LCMS (m/z)=564.2 [M+H], Tr=1.96 min.
Example 74
##STR00374##
Compound 57 (20 mg, 0.036 mmol) was dissolved in methanol (5 mL)
and added catalytic amount of 10% Pd on carbon. The reaction
mixture was stirred under atmosphere hydrogen for 2 h. The catalyst
was removed by filtration and the eluent was concentrated under
reduced pressure. Purification by reverse phase preparative HPLC
gave the title compound (8.4 mg, 42% yield) as a white powder.
.sup.1H NMR (300 MHz, CD.sub.3OD): .delta.9.61 (s, 1H), 8.42 (d,
J=9.0 Hz, 1H), 7.99 (s, 1H), 7.89 (d, J=9.0 Hz, 1H), 7.83 (br s,
1H), 7.29 (d, J=8.7 Hz, 1H), 6.27-6.21 (m, 1H), 5.92-5.82 (m, 1H),
3.96-3.5 (m, 4H), 3.28-3.19 (m, 1H), 3.08-2.98 (m, 1H), 2.30-2.20
(m, 1H), 2.10-1.90 (m, 4H), 1.78-1.72 (m, 5H), 1.56-1.51 (m, 6H),
1.21 (s, 3H), 0.89 (d, J=7.2 Hz, 6H). LCMS (m/z): 552.3 [M+H],
Tr=1.74 min.
Example 75
##STR00375##
A solution of ethyl acetoacetate (20 g, 19.4 mL, 0.154 mol) in a
mixture of dioxane (120 mL) and aqueous formaldehyde (37% solution
in water, 57.7 mL, 0.77 mol) was stirred at RT. Triethylamine (1.0
M in tetrahydrofuran, 7.7 mL, 7.7 mmol) was added and the reaction
mixture was heated at 60.degree. C. for 20 h and then heated at
100.degree. C. for 4 h. The reaction mixture was cooled to RT and
was poured into water (1500 mL). The aqueous solution was washed
with toluene. The aqueous layer was concentrated to .about.50% of
the initial volume and extracted with ethyl acetate. The organic
extracts were combined and evaporated. The residue was purified by
silica gel chromatography using a gradient of iso-hexane to
iso-hexane/ethyl acetate 1:1 followed by silica gel chromatography
using a gradient of iso-hexane to iso-hexane/ethyl acetate 3:2 to
afford the title compound (2.33 g, 8%) as a yellow oil.
##STR00376##
A solution of 75a (2.10 g, 11 mmol), 2,2-dimethoxypropane (13.5 mL,
110 mmol) and 4-toluenesulfonic acid hydrate (209 mg, 1.1 mmol) in
acetone (8 mL) was stirred at RT for 18 h. Saturated sodium
hydrogen carbonate solution was added and the mixture was extracted
with ethyl acetate. The organic extracts were combined, washed with
water and brine. The organic layer was filtered through a
hydrophobic frit and the filtrate was evaporated. The residue was
purified by silica gel chromatography using a gradient of
iso-hexane/ethyl acetate 1:5 to 3:7 to afford the title compound
(1.88 g, 74%) as a colorless oil.
##STR00377##
A solution of N,N-diisopropylethylamine (0.55 mL, 3.9 mmol) in
tetrahydrofuran (20 mL) was stirred at -78.degree. C. under
nitrogen. n-Butyl lithium (1.6 M in hexane, 2.25 mL, 3.6 mmol) was
added dropwise and the reaction mixture was warmed to 0.degree. C.
The reaction mixture was stirred at 0.degree. C. for 5 min and then
cooled to -78.degree. C. A solution of 75b (690 mg, 3.0 mmol) in
tetrahydrofuran (3 mL) was added dropwise and the reaction mixture
was stirred at -78.degree. C. for 15 min. A solution of
N-phenyl-(bistrifluoromethanesulfonamide) (1.18 g, 3.3 mmol) in
tetrahydrofuran (10 mL) was added dropwise over 5 min and the
reaction mixture was stirred at -78.degree. C. for 15 min. The
cooling bath was removed and the reaction mixture was warmed to RT
and then stirred at RT for 90 min. The solvent was evaporated and
diethyl ether (30 mL) was added. The solution was cooled to
5.degree. C. and was washed with cold 1 M sodium hydroxide solution
(3.times.30 mL). The organic layer was separated, washed with
brine, dried over anhydrous sodium sulfate, filtered and the
solvent evaporated to afford the title compound (960 mg, 88%) as a
yellow oil which was used directly in the next step.
##STR00378##
A solution of 75c (470 mg, 1.3 mmol) and tri-n-butylamine (721 mg,
0.93 mL, 3.9 mmol) in N,N-dimethylformamide (3 mL) was stirred at
RT under nitrogen. Bis(triphenylphosphine)palladium(II) dichloride
(45 mg, 0.065 mmol) and formic acid (120 mg, 0.1 mL, 2.6 mmol) was
added and the reaction mixture was heated at 60.degree. C. for 90
min. The reaction mixture was cooled to RT and ethyl acetate and
water was added. The organic extract was separated, washed with
water (.times.5), and brine. The organic solution was filtered
through a hydrophobic frit and the filtrate was evaporated. The
residue was purified by silica gel chromatography using
iso-hexane/ethyl acetate 1:9 to afford the title compound (1.88 g,
74%) as a colorless oil.
##STR00379##
A solution of 75d (150 mg, 0.7 mmol) in tetrahydrofuran (3 mL) was
stirred at 5.degree. C. under nitrogen. A solution of lithium
hydroxide monohydrate (59 mg, 1.4 mmol) in water (1 mL) was added
and the reaction mixture was stirred at 5.degree. C. for 30 min and
then at RT for 5 h. Methanol (0.5 mL) was added to give a clear
solution and the reaction mixture was stirred at RT for 22 h. The
solvent was evaporated. Water (2 mL) was added to the residue and
the solution was acidified to pH 2 with 2 M hydrochloric acid.
Brine was added and the mixture was extracted with ethyl acetate.
The organic extracts were combined, washed with brine. The organic
extract was separated, washed with water (.times.5), and brine. The
organic solution was filtered through a hydrophobic frit and the
filtrate was evaporated to afford the title compound (117 mg, 90%)
as a colorless oil.
##STR00380##
A mixture of 75e (96 mg, 0.5 mmol) and acetic acid
(R)-1-(7-bromo-quinolin-2-yl)-ethyl ester (147 mg, 0.5 mmol) in
acetonitrile (4 mL) was stirred at RT. Tri(o-tolyl)phosphine (46
mg, 0.15 mmol), palladium(II) acetate (17 mg, 0.075 mmol) and
triethylamine (101 mg, 0.14 mL, 1.0 mmol) was added and the
reaction mixture was heated in a microwave reactor at 100.degree.
C. for 20 min. The solvent was evaporated. Water and ethyl acetate
was added and the mixture was acidified to pH 3-4 with 2 M
hydrochloric acid. The mixture was extracted with ethyl acetate and
the organic extracts were combined and washed with brine. The
organic solution was filtered through a hydrophobic frit and the
filtrate was evaporated. The residue was purified by silica gel
chromatography using iso-hexane/ethyl acetate 1:1 to ethyl acetate
to ethyl acetate/methanol 5:1. The residue was co-evaporated with
ethyl acetate and then dichloromethane and dried to afford the
title compound (134 mg, 67%) as a yellow gum.
##STR00381##
A solution of
(S)-1-[(S)-2-((S)-2-tert-butoxycarbonylamino-3-methyl-butyrylamino)-propi-
onyl]-hexahydro-pyridazine-3-carboxylic acid methyl ester (150 mg,
0.36 mmol) in dichloromethane (5 mL) was stirred at 0.degree. C.
under nitrogen. Trimethylsilyl trifluoromethanesulfonate (160 mg,
0.13 mL, 0.72 mmol) was added and the reaction mixture was stirred
at 0.degree. C. for 1 hour. N,N-Diisopropylethylamine (186 mg, 0.25
mL, 1.44 mmol) was added and the solvent was evaporated to afford
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid methyl ester (0.36 mmol) as a white solid
which was used without further purification. A solution of 75f (134
mg, 0.33 mmol) in acetonitrile was stirred at 0.degree. C. under
nitrogen. A solution of
(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyr-
idazine-3-carboxylic acid methyl ester (0.36 mmol) and
N,N-diisopropylethylamine (129 mg, 0.17 mL, 1.0 mmol) in
acetonitrile (5 mL) was added followed by
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (175 mg, 0.46 mmol) and the
reaction mixture was stirred at 0.degree. C. for 10 min and then at
RT for 1 hour. The solvent was evaporated and the residue was
partioned between ethyl acetate and water. The organic extracts
were combined and washed with brine. The organic solution was
filtered through a hydrophobic frit and the filtrate was
evaporated. The residue was purified by silica gel chromatography
using iso-hexane/ethyl acetate 1:5 to ethyl acetate to afford the
title compound (165 mg, 72%) as a yellow solid.
##STR00382##
A solution of 75g (35 mg, 0.05 mmol) in tetrahydrofuran (0.5 mL)
was stirred at 5.degree. C. under nitrogen. A solution of lithium
hydroxide monohydrate (8.4 mg, 0.2 mmol) in water (0.5 mL) was
added followed by methanol (0.5 mL) and the reaction mixture was
stirred at 5.degree. C. for 1 hour. 1 M hydrochloric acid (0.2 mL)
was added and the solvent was evaporated. The residue was
co-evaporated with methanol/toluene (1:1, .times.2) followed by
toluene (.times.2). The residue was triturated with diethyl ether
(.times.3) and dried to afford the title compound (0.05 mmol) as a
pale yellow solid which was used crude in the next reaction.
##STR00383##
A solution of 2-methyl-6-nitrobenzoic anhydride (86 mg, 0.25 mmol)
and 4-dimethylaminopyridine (46 mg, 0.38 mmol) in
1,2-dichloroethane (16 mL) was stirred at RT under nitrogen. A
molecular sieves (400 mg) was added and the suspension was heated
at 50.degree. C. A solution of crude 75h (0.05 mmol) in
N,N-dimethylformamide (2 mL) was added dropwise over 4 h and the
reaction mixture was stirred at 50.degree. C. for an additional 1
hour. The reaction mixture was cooled to RT and the mixture was
filtered through celite. The filter pad was washed with ethyl
acetate and the filtrate was evaporated. The residue was diluted
with ethyl acetate and the solution was washed with brine
(.times.3). The organic layer was separated and evaporated. The
organic solution was filtered through a hydrophobic frit and the
filtrate was evaporated. The residue was purified by preparative
HPLC to afford the title compound (4.2 mg, 13%) as a white solid.
.sup.1H NMR (300 MHz, CD.sub.3OD) 1.00 (d, J=6.7 Hz, 3H), 1.01 (d,
J=6.6 Hz, 3H), 1.37 (s, 3H), 1.47 (s, 3H), 1.63 (d, J=7.1 Hz, 3H),
1.64-1.70 (m, 2H), 1.73 (d, J=6.9 Hz, 3H), 1.85-2.10 (m, 3H),
2.70-2.80 (m, 1H), 3.78-3.87 (m, 1H), 3.87 (d, J=11.1 Hz, 1H),
4.03-4.07 (m, 1H), 4.26-4.46 (m, 5H), 5.72 (q, J=7.1 Hz, 1H), 5.93
(q, J=6.8 Hz, 1H), 6.38 (s, 2H), 7.42 (d, J=8.5 Hz, 1H), 7.61 (br
s, 1H), 7.78-7.86 (m, 2H), 8.22 (d, J=8.5 Hz, 1H). LCMS (m/z) 622.2
[M+H], Tr=2.24 min.
Example 76
##STR00384##
A suspension of crude 75h (200 mg, 0.25 mmol) in tetrahydrofuran
(100 mL) was stirred at RT under nitrogen.
N,N-diisopropylethylamine (161 mg, 0.22 mL, 1.25 mmol) and
4-dimethylaminopyridine (15 mg, 0.125 mmol) was added and the
suspension was stirred for 5 min.
2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (133 mg, 0.35 mmol) was added and
the reaction mixture was stirred at RT for 2 h. Additional
2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate methanaminium (57 mg, 0.15 mmol) was added and
the reaction mixture was stirred at RT for 1 hour. The solvent was
evaporated and the residue was diluted with ethyl acetate and 2 M
hydrochloric acid. The organic solution was separated and washed
with water, saturated sodium hydrogen carbonate solution, and
brine. The organic solution was filtered through a hydrophobic frit
and the filtrate was evaporated. The residue was purified by silica
gel chromatography using iso-hexane/ethyl acetate 7:3 to ethyl
acetate to ethyl acetate/methanol 9:1. The residue was purified by
preparative HPLC to afford the title compound (20 mg, 14%) as a
white solid. .sup.1H NMR (500 MHz, CD.sub.3OD) 1.00 (d, J=6.5 Hz,
3H), 1.01 (d, J=6.5 Hz, 3H), 1.68 (d, J=7.1 Hz, 3H), 1.73 (d, J=6.7
Hz, 3H), 1.74-2.05 (m, 5H), 2.72-2.80 (m, 1H), 3.70 (d, J=11.1 Hz,
1H), 3.81-4.07 (m, 4H), 4.35-4.47 (m, 3H), 5.77 (q, J=7.1 Hz, 1H),
5.92 (q, J=6.8 Hz, 1H), 6.43 (s, 2H), 7.40 (d, J=8.5 Hz, 1H), 7.63
(s, 1H), 7.82 (s, 2H), 8.21 (d, J=8.5 Hz, 1H). LCMS (m/z) 582.2
[M+H], Tr=1.66 min.
Biological Examples: Inhibition of Peptidyl-Prolyl Isomerase
(PPlase) Activity
The PPlase assay was based on the procedure reported by Janowski et
al. (Anal. Biochem. 1997, 252, 299). Assay buffer (1980 L of a
solution containing 35 mM HEPES pH 7.8, 50M DTT, and 0.01% N P40)
was pre-equilibrated to 10.degree. C. in a quartz cuvette equipped
with an overhead stirrer. To this solution was added 10 L of
compound in DMSO (final concentration: 0.5% DMSO), followed by 5 L
of a 2M stock solution of cyclophilin A (final concentration: 5
nM). The reaction was initiated with the addition of 5 L of 40 mM
of the tetrapeptide Succ-AAPF-pNA (100M final concentration)
dissolved in a solution of 0.5M LiCl in trifluoroethanol. Upon the
initiation of the reaction, the absorbance of the peptide substrate
was monitored at 330 nm for five minutes using a Beckman Coulter
DU800 spectrophotometer. Progress curves were fit with a
single-exponential decay model to calculate rates. The IC.sub.50
values were calculated with a four-parameter logistic fit using
GraphPad Prism software.
Cyclophilin A TR-FRET Competitive Binding Assay
Inhibitor potency was measured using a competitive binding assay
with a time-resolved fluorescence resonance energy transfer
(TR-FRET) readout. To a reaction buffer consisting of 35 mM HEPES
pH 7.8, 100 mM NaCl, 0.01% NP-40 (Pierce), 1 mM DTT, and 1% DMSO
were added the following: 5 nM of cyclophilin A modified at the
N-terminus with an 8.times. histidine affinity tag (CypA); 150 nM
of cyclosporin A modified with a linker attached to a Cy5
fluorophore (CsA-Cy5); 1 nM Eu-labeled anti-(6.times.His) antibody
(Perkin-Elmer); and test compound at one of various concentrations.
The total volume of the assay solution was 100 L. After a two-hour
incubation, the TR-FRET was measured using a Perkin Elmer Envision
plate reader (excitation at 340 nm, emission measured at 590 nm and
665 nm). The signal was calculated as the ratio of the emission at
665 nm to that at 590 nm. An IC.sub.50 value was calculated using a
four-parameter logistic fit.
When tested, certain compounds of this invention were found to
inhibit cyclophilin binding as listed in Table 1 below. The
IC.sub.50's are presented as ranges wherein A is 100 nM, B is 101
to 1000 nM and C is 1001 to 10,000 nM.
Antiviral Activity
The antiviral activity of a compound can be measured using standard
screening protocols: for example, cell-based Flavivirus
immunodetection assay and cell-based Flavivirus cytopathic effect
assay as described in U.S. Patent Publication Number
US/20130022573, which is hereby incorporated by reference in its
entirety.
One aspect of the invention relates to methods of inhibiting viral
infections, comprising the step of treating a sample or subject
suspected of needing such inhibition with a composition of the
invention. The antiviral activity of a compound of the invention
can be measured using standard screening protocols that are
known.
The anti-HCV activity of the compounds of this invention was tested
in a human hepatoma Huh-7 cell line harboring a HCV replicon. The
assay comprised the following steps:
Step 1 (Compound Preparation and Serial Dilution):
Serial dilution was performed in 100% DMSO in a 384-well plate. A
solution containing a compound at 225-fold concentration of the
starting final serial dilution concentration was prepared in 100%
DMSO and 15 .mu.L added to the pre-specified wells in column 3 or
13 of a polypropylene 384-well plate. The rest of the 384-well
plate was filled with 10 .mu.L 100% DMSO except for columns 23 and
24, where 10 .mu.L of 500 .mu.M a HCV protease inhibitor (ITMN-191)
in 100% DMSO was added. The HCV protease inhibitor was used a
control of 100% inhibition of HCV replication. The plate was then
placed on a Biomek FX Workstation to start the serial dilution. The
serial dilution was performed for ten cycles of 3-fold dilution
from column 3 to 12 or from column 13 to 22.
Step 2 (Cell Culture Plate Preparation and Compound Addition):
To each well of a black polypropylene 384-well plate, 90 .mu.L of
cell media containing 1600 suspended Huh-7 HCV replicon cells was
added with a Biotek uFlow Workstation. A volume of 0.4 .mu.L of the
compound solution was transferred from the serial dilution plate to
the cell culture plate on a Biomek FX Workstation. The DMSO
concentration in the final assay condition was 0.44%. The plates
were incubated for 3 days at 37.degree. C. with 5% CO.sub.2 and 85%
humidity.
Step 3 (Detection of Cytotoxicity and Inhibition of Viral
Replication):
a) Assessment of cytotoxicity: The media in the 384-well cell
culture plate was aspirated with a Biotek EL405 plate-washer. A
volume of 50 .mu.L of a solution containing 400 nM Calcein AM in
100% PBS was added to each well of the plate with a Biotek uFlow
Workstation. The plate was incubated for 30 minutes at RT before
the fluorescence signal (emission 490 nm, exitation 520 nm) was
measured with a Perkin Elmer Envision Plate Reader.
b) Assessment of inhibition of viral replication: The calcein-PBS
solution in the 384-well cell culture plate was aspirated with a
Biotek EL405 plate-washer. A volume of 20 .mu.L of Dual-Glo
luciferase buffer (Promega, Dual-Glo Luciferase Assay Reagent, cat.
#E298B) was added to each well of the plate with a Biotek uFlow
Workstation. The plate was incubated for 10 minutes at RT. A volume
of 20 .mu.L of a solution containing 1:100 mixture of Dual-Glo Stop
& Glo substrate (Promega, Dual-Glo Luciferase Assay Reagent,
cat. #E313B) and Dual-Glo Stop & Glo buffer (Promega, Dual-Glo
Luciferase Assay Reagent, cat. #E314B) was then added to each well
of the plate with a Biotek uFlow Workstation. The plate was
incubated at RT for 10 minutes before the luminescence signal was
measured with a Perkin Elmer Envision Plate Reader.
Step 4 (Calculation):
The percent cytotoxicity was determined by calcein AM conversion to
fluorescent product. The average fluorescent signal from the DMSO
control wells were defined as 100% nontoxic. The individual
fluorescent signal from testing compound treated well was divided
by the average signal from DMSO control wells and then multiplied
by 100% to get the percent viability. The percent anti-HCV
replication activity was determined by the luminescence signal from
the testing well compared to DMSO controls wells. The background
signal was determined by the average luminescence signal from the
HCV protease inhibitor treated wells and was subtracted from the
signal from the testing wells as well as the DMSO control wells.
Following 3-fold serial dilutions, the EC.sub.50 and CC.sub.50
values were calculated by fitting % inhibition at each
concentration to the following equation: %
inhibition=100%/[(EC.sub.50/[I]).sup.b+1]
where b is Hill's coefficient. See, for reference, Hill, A. V., The
Possible Effects of the Aggregation of the Molecules of H.ae
butted.moglobin on its Dissociation Curves, J. Physiol. 40: iv-vii.
(1910). % inhibition values at a specific concentration, for
example 2 .mu.M, can also be derived from the formula above.
When tested, certain compounds of this invention were found to
inhibit viral replication as listed in Table 1. The EC.sub.50's are
presented as a % inhibition.
TABLE-US-00007 TABLE 1 Example TR- Replicon 1a No. FRET %
inhibition at 1 .mu.m 1 A 32 2 B 63 3 A 44 4 B 58 5 A 80 6 A 86 7 A
94 8 B 37 9 A 98 10 B 28 11 A 6 12 C 0 13 A 87 14 A 94 15 A 99 16 B
94 17 A 100 18 A 94 19 A 97 20 A 99 21 A 93 22 A 100 23 A 99 24 A
99 25 B 71 26 A 99 27 A 100 28 B 56 29 C 28 30 A 73 31 A 98 32 A 71
33 C 14 34 A 92 35 A 94 36 A 73 37 A 76 38 A 98 39 A 98 40 A 96 41
A 96 42 B 16 43 A 97 44 A 78 45 A 80 46 A 90 47 B 9 48 A 40 49 A 85
50 B 12 51 A 83 52 A 98 53 B 89 54 A 98 55 B 76 56 B 47 57 A 100 58
A 99 59 A 100 60 B 65 61 A 72 62 B 25 63 B 80 64 B 35 65 A 92 66 A
80 67 A 97 68 A 91 69 C 19 70 A 95 71 B -- 72 B 0 73 A 94 74 A 95
75 A 100 76 A 63
The specific pharmacological and biochemical responses observed may
vary according to and depending on the particular active compound
selected or whether there are present pharmaceutical carriers, as
well as the type of formulation and mode of administration
employed, and such expected variations or differences in the
results are contemplated in accordance with practice of the present
invention.
Although specific embodiments of the present invention are herein
illustrated and described in detail, the invention is not limited
thereto. The above detailed descriptions are provided as exemplary
of the present invention and should not be construed as
constituting any limitation of the invention.
* * * * *
References